Tetrahydropyranyl cyclopentyl benzylamide modulators of chemokine receptor activity

ABSTRACT

The present invention is directed to compounds of the formula (I) (wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 27 , R 28 , R 29 , R 30 , R 31 , X, m, n and the dashed line are defined herein) which are useful as modulators of chemokine receptor activity. In particular, these compounds are useful as modulators of the chemokine receptor CCR-2.

BACKGROUND OF THE INVENTION

The chemokines are a family of small (70-120 amino acids),proinflammatory cytokines, with potent chemotactic activities.Chemokines are chemotactic cytokines that are released by a wide varietyof cells to attract various cells, such as monocytes, macrophages, Tcells, eosinophils, basophils and neutrophils to sites of inflammation(reviewed in Schall, Cytokine, 3, 165-183 (1991) and Murphy, Rev.Inmun., 12, 593-633 (1994)). These molecules were originally defined byfour conserved cysteines and divided into two subfamilies based on thearrangement of the first cysteine pair. In the CXC-chemokine family,which includes IL-8, GROα, NAP-2 and IP-10, these two cysteines areseparated by a single amino acid, while in the CC-chemokine family,which includes RANTES, MCP-1, MCP-2, MCP-3, MIP-1α, MIP-1β and eotaxin,these two residues are adjacent.

The α-chemokines, such as interleukin-8 (IL-8), neutrophil-activatingprotein-2 (NAP-2) and melanoma growth stimulatory activity protein(MGSA) are chemotactic primarily for neutrophils, whereas β-chemokines,such as RANTES, MIP-1α, MIP-1β, monocyte chemotactic protein-1 (MCP-1),MCP-2, MCP-3 and eotaxin are chemotactic for macrophages, monocytes,T-cells, eosinophils and basophils (Deng, et al., Nature, 381, 661-666(1996)).

The chemokines are secreted by a wide variety of cell types and bind tospecific G-protein coupled receptors (GPCRs) (reviewed in Horuk, TrendsPharm. Sci., 15, 159-165 (1994)) present on leukocytes and other cells.These chemokine receptors form a sub-family of GPCRs, which, at present,consists of fifteen characterized members and a number of orphans.Unlike receptors for promiscuous chemoattractants such as C5a, fMLP,PAF, and LTB4, chemokine receptors are more selectively expressed onsubsets of leukocytes. Thus, generation of specific chemokines providesa mechanism for recruitment of particular leukocyte subsets.

On binding their cognate ligands, chemokine receptors transduce anintracellular signal though the associated trimeric G protein, resultingin a rapid increase in intracellular calcium concentration. There are atleast seven human chemokine receptors that bind or respond toβ-chemokines with the following characteristic pattern: CCR-1 (or“CKR-1” or “CC-CKR-1”) [MP-1α, MIP-1β, MCP-3, RANTES] (Ben-Barruch, etal., J. Biol. Chem., 270, 22123-22128 (1995); Beote, et al, Cell, 72,415-425 (1993)); CCR-2A and CCR-2B (or “CKR-2A”/“CKR-2A” or“CC-CKR-2A”/“CC-CKR-2A”) [MCP-1, MCP-2, MCP-3, MCP-4]; CCR-3 (or “CKR-3”or “CC-CKR-3”) [Eotaxin, Eotaxin 2, RANTES, MCP-2, MCP-3] (Rollins, etal., Blood, 90,908-928 (1997)); CCR-4 (or “CKR-4” or “CC-CKR-4”) [MP-1α,RANTES, MCP-1] (Rollins, et al., Blood, 90, 908-928 (1997)); CCR-5 (or“CKR-5” or “CC-CKR-5”) [MIP-1α, RANTES, MIP-1β] (Sanson, et al.,Biochemistry, 35, 3362-3367 (1996)); and the Duffy blood-group antigen[RANTES, MCP-1] (Chaudhun, et al., J. Biol. Chem., 269, 7835-7838(1994)). The β-chemokines include eotaxin, MIP (“macrophage inflammatoryprotein”), MCP (“monocyte chemoattractant protein”) and RANTES(“regulation-upon-activation, normal T expressed and secreted”) amongother chemokines.

Chemokine receptors, such as CCR-1, CCR-2, CCR-2A, CCR-2B, CCR-3, CCR-4,CCR-5, CXCR-3, CXCR-4, have been implicated as being important mediatorsof inflammatory and immunoregulatory disorders and diseases, includingasthma, rhinitis and allergic diseases, as well as autoimmunepathologies such as rheumatoid arthritis and atherosclerosis. Humans whoare homozygous for the 32-basepair deletion in the CCR-5 gene appear tohave less susceptibility to rheumatoid arthritis (Gomez, et al.,Arthritis & Rheumatism, 42, 989-992 (1999)). A review of the role ofeosinophils in allergic inflammation is provided by Kita, H., et al., J.Exp. Med. 183, 2421-2426 (1996). A general review of the role ofchemokines in allergic inflammation is provided by Lustger, A. D., NewEngland J. Med., 338(7), 426-445 (1998).

A subset of chemokines are potent chemoattractants for monocytes andmacrophages. The best characterized of these is MCP-1 (monocytechemoattractant protein-1), whose primary receptor is CCR2. MCP-1 isproduced in a variety of cell types in response to inflammatory stimuliin various species, including rodents and humans, and stimulateschemotaxis in monocytes and a subset of lymphocytes. In particular,MCP-1 production correlates with monocyte and macrophage infiltration atinflammatory sites. Deletion of either MCP-1 or CCR2 by homologousrecombination in mice results in marked attenuation of monocyterecruitment in response to thioglycollate injection and Listeriamonocytogenes infection (Lu et al., J. Exp. Med., 187, 601-608 (1998);Kurihara et al. J. Exp. Med., 186, 1757-1762 (1997); Boring et al. J.Clin. Invest., 100, 2552-2561 (1997); Kuziel et al. Proc. Natl. Acad.Sci., 94, 12053-12058 (1997)). Furthermore, these animals show reducedmonocyte infiltration into granulomatous lesions induced by theinjection of schistosomal or mycobacterial antigens (Boring et al. J.Clin. Invest., 100, 2552-2561 (1997); Warmington et al. Am J. Path.,154, 1407-1416 (1999)). These data suggest that MCP-1-induced CCR2activation plays a major role in monocyte recruitment to inflammatorysites, and that antagonism of this activity will produce a sufficientsuppression of the immune response to produce therapeutic benefits inimmunoinflammatory and autoimmune diseases

Accordingly, agents which modulate chemokine receptors such as the CCR-2receptor would be useful in such disorders and diseases.

In addition, the recruitment of monocytes to inflammatory lesions in thevascular wall is a major component of the pathogenesis of atherogenicplaque formation. MCP-1 is produced and secreted by endothelial cellsand intimal smooth muscle cells after injury to the vascular wall inhypercholesterolemic conditions. Monocytes recruited to the site ofinjury infiltrate the vascular wall and differentiate to foam cells inresponse to the released MCP-1. Several groups have now demonstratedthat aortic lesion size, macrophage content and necrosis are attenuatedin MCP-1 −/− or CCR2 −/− mice backcrossed to APO-E −/−, LDL-R −/− or ApoB transgenic mice maintained on high fat diets (Boring et al. Nature,394, 894897 (1998); Gosling et al. J. Clin. Invest., 103, 773-778(1999)). Thus, CCR2 antagonists may inhibit atherosclerotic lesionformation and pathological progression by impairing monocyte recruitmentand differentiation in the arterial wall.

SUMMARY OF THE INVENTION

The present invention is further directed to compounds which aremodulators of chemokine receptor activity and are useful in theprevention or treatment of certain inflammatory and immunoregulatorydisorders and diseases, allergic diseases, atopic conditions includingallergic rhinitis, dermatitis, conjunctivitis, and asthma, as well asautoimmune pathologies such as rheumatoid arthritis and atherosclerosis.The invention is also directed to pharmaceutical compositions comprisingthese compounds and the use of these compounds and compositions in theprevention or treatment of such diseases in which chemokine receptorsare involved.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to compounds of the formula I:

-   X is selected from the group consisting of:    -   —O—, —NR²⁰—, —S—, —SO—, —SO₂—, and —CR²¹R²²—, —NSO₂R²⁰—,        —NCOR²⁰—, —NCO₂R²⁰—, —CR²¹CO₂R²⁰—, —CR²¹OCOR²⁰—, —CO—,        C(CH₃)₂—O—,    -   where R²⁰ is selected from: hydrogen, C₁₋₆ alkyl, benzyl,        phenyl, C₃₋₆ cycloalkyl where the alkyl, phenyl, benzyl, and        cycloalkyl groups can be unsubstituted or substituted with 1-3        substituents where the substituents are independently selected        from: halo, hydroxy, C₁₋₃alkyl, C₁₋₃alkoxy, —CO₂H, —CO₂—C₁₋₆        alkyl, and trifluoromethyl,    -   where R²¹ and R²² are independently selected from: hydrogen,        hydroxy, C₁₋₆ alkyl, —O—C₁₋₆alkyl, benzyl, phenyl, C₃₋₆        cycloalkyl where the alkyl, phenyl, benzyl, and cycloalkyl        groups can be unsubstituted or substituted with 1-3 substituents        where the substituents are independently selected from: halo,        hydroxy, C₁₋₃alkyl, C₁₋₃alkoxy, —CO₂H, —CO₂—C₁₋₆ alkyl, and        trifluoromethyl;-   R¹ is selected from:    -   —C₁₋₆alkyl, —C₀₋₆alkyl-O—C₁₋₆alkyl, —C₀₋₆alkyl-S—C₁₋₆alkyl,        —C₀₋₆alkyl-SO₁₋₂—C₁₋₆alkyl, —C₀₋₆alkyl-SO₂—NR²⁶—C₁₋₆alkyl,        —(C₀₋₆alkyl)-(C₃₋₇cycloalkyl)-(C₀₋₆alkyl), hydroxy, —CO₂R²⁰,        heterocycle, —CN, —NR²⁰R²⁶, —NR²⁶SO₂R²⁰, —NR²⁶COR²¹, —OCOR²⁰,        and phenyl, where R²⁶ is selected from: hydrogen, C₁₋₆ alkyl,        benzyl, phenyl, C₃₋₆ cycloalkyl where the alkyl, phenyl, benzyl,        and cycloalkyl groups can be unsubstituted or substituted with        1-3 substituents where the substituents are independently        selected from: halo, hydroxy, C₁₋₃alkyl, C₁₋₃alkoxy, —CO₂H,        —CO₂—C₁₋₁₆ alkyl, and trifluoromethyl    -   where the allyl and the cycloalkyl are unsubstituted or        substituted with 1-7 substituents where the substituents are        independently selected from: halo, hydroxy, —O—C₁₋₃alkyl,        trifluoromethyl, C₁₋₃alkyl, —O—C₁₋₃alkyl, —CO₂R²⁰, —SO₂R²⁰,        —NHCOCH₃, —NHSO₂CH₃, -heterocycle, ═O, —CN,    -   and where the phenyl and heterocycle are unsubstituted or        substituted with 1-3 substituents where the substituents are        independently selected from: halo, hydroxy, C₁₋₃alkyl,        C₁₋₃alkoxy and trifluoromethyl;-   R² is selected from: hydrogen, C₁₋₆alkyl, trifluoromethyl,    trifluoromethoxy, chloro, bromo, and phenyl;-   R³ is selected from: hydrogen, hydroxy, halo, C₁₋₆alkyl,    —O—C₁₋₆alkyl, —NR²⁰R²¹, —NR²⁰CO₂R²¹, —NR²⁰CONR²⁰R²¹,    —NR²⁰—SO₂—NR²⁰R²¹, —NR²⁰—SO₂—R²¹, heterocycle, —CN, —CONR²⁰R²¹,    —CO₂R²⁰, —NO₂, —S—R²⁰, —SO—R²⁰, —SO₂—R²⁰, and —SO₂—NR²⁰R²¹;-   R⁴ is selected from: hydrogen, C₁₋₆alkyl, trifluoromethyl,    trifluoromethoxy, chloro, bromo, and phenyl;-   R⁵ is selected from: C₁₋₆alkyl substituted with 1-6 fluoro and    optionally substituted with hydroxyl, —O—C₁₋₆alkyl substituted with    1-6 fluoro, —CO—C₁₋₆alkyl substituted with 1-6 fluoro, —S—C₁₋₆alkyl,    -pyridyl, fluoro, chloro, bromo, and phenyl;-   R⁶ is selected from: hydrogen, C₁₋₆alkyl, trifluoromethyl,    trifluoromethoxy, chloro, bromo, and phenyl;-   R⁷ is selected from: hydrogen, C₁₋₆alkyl, and trifluoromethyl;-   R⁸ is selected from: hydrogen, C₁₋₆alkyl, where alkyl may be    unsubstituted or substituted with 1-6 substituents where the    substituents are chosen from the group: fluoro, C₁₋₃alkoxy, hydroxy,    —CO₂R²⁰, fluoro, —O—C₁₋₃alkyl, where alkyl may be unsubstituted or    substituted with 1-3 fluoro, and C₃₋₆ cycloalkyl, —O—C₃-cycloalkyl,    hydroxy, —CO₂R²⁰, —OCOR²⁰, phenyl,    -   or R⁷ and R⁸ may be joined together via a C₂₋₄alkyl or a        C₀₋₂alkyl-O—C₁₋₃alkyl chain to form a 5-7 membered ring;-   R⁹ is selected from: hydrogen, C₁₋₆alkyl, where alkyl may be    unsubstituted or substituted with 1-6 substituents where the    substituents are chosen from the group: fluoro, C₁₋₃alkoxy, hydroxy,    —CO₂R²⁰, —CO₂R²⁰, hydroxy, and —O—C₁₋₆alkyl, where alkyl may be    unsubstituted or substituted with 1-6 substituents where the    substituents are chosen from the group: fluoro, C₁₋₃alkoxy, hydroxy,    —CO₂R²⁰,    -   or R⁸ and R⁹ may be joined together by a C₁₋₄alkyl chain or a        C₀₋₃alkyl-O—C₀₋₃alkyl chain to form a 3-6 membered ring;        R¹⁰ is selected from: hydrogen, and C₁₋₆alkyl, where alkyl may        be unsubstituted or substituted with 1-6 fluoro, fluoro,        —O—C₃Cycloalkyl, and —O—C₁₋₃alkyl, where alkyl may be        unsubstituted or substituted with 1-6 fluoro,    -   or R⁸ and R¹⁰ may be joined together by a C₁₋₃alkyl chain or a        single bond to form a 3-6 membered ring; where the alkyl are        unsubstituted or substituted with 1-3 substituents where the        substiuents are independently selected from: halo, hydroxy,        —CO₂R²⁰, C₁₋₃alkyl, and C₁₋₃alkoxy,    -   or R⁸ and R¹⁰ may be joined together by a C₁₋₂alkyl-O—C₁₋₂alkyl        chain to form a 6-8 membered ring, where the alkyl are        unsubstituted or substituted with 1-3 substituents where the        substiuents are independently selected from: halo, hydroxy,        —CO₂R²⁰, C₁₋₃alkyl, and C₁₋₃alkoxy,    -   or R⁸ and R¹⁰ may be joined together by a —O—C₁₋₂alkyl-O— chain        to form a 6-7 membered ring, where the alkyl are unsubstituted        or substituted with 1-3 substituents where the substiuents are        independently selected from: halo, hydroxy, —CO₂R²⁰, C₁₋₃alkyl,        and C₁₋₃alkoxy;-   R¹¹ is selected from: hydrogen, C₁₋₆alkyl, and trifluoromethyl;-   R²⁷ and R²⁸ are independently selected from: ═O, where R²⁷, R²⁸, or    both, is oxygen and is connected via a double bond, hydrogen,    phenyl, and C₁₋₆alkyl which may be substituted or unsubstituted with    1-6 of the following substituents: —COR¹¹, hydroxy, fluoro, chloro,    —O—C₁₋₃alkyl;-   R²⁹, R³⁰, and R³¹ are independently selected from: hydrogen, methyl,    hydroxyl, trifluoromethyl, methoxy, and trifluoromethoxy;-   or R²⁹ and R⁹ are connected by a C₁₋₃alkyl bridge;-   m is selected from 0, 1, and 2;-   n is selected from 0, 1 and 2;-   the dashed line represents a single or a double bond;    and pharmaceutically acceptable salts thereof and individual    diastereomers thereof.

Preferred compounds of the present invention include those of formulaIa:

wherein R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰ and X are definedherein;and pharmaceutically acceptable salts and individual diastereomersthereof.

Preferred compounds of the present invention also include those offormula Ib:

wherein R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, and R¹⁰ are defined herein,and Y is selected from the group consisting of: —O—, —CH₂—, —S—, —SO—,and —SO₂—;and pharmaceutically acceptable salts and individual diastereomersthereof.

More preferred compounds of the present invention also include those offormula Ic:

wherein R¹, R², R³, R⁵ and R⁸ are defined herein.

More preferred compounds of the present invention also include those offormula Id:

wherein:R¹ is selected from: C₁₋₆alkyl, C₁₋₆alkyl-hydroxy, and C₁₋₆alkylsubstituted with 1-6 fluoro;R³ is selected from: C₁₋₆alkyl unsubstituted or substituted with 1-6fluoro, fluoro; chloro, bromo, and phenyl;R⁵ is selected from: C₁₋₆alkyl unsubstituted or substituted with 1-6fluoro, fluoro; chloro, bromo, and phenyl;R⁸ is selected from: hydrogen, C₁₋₆alkyl, C₁₋₆alkyl-hydroxy, andC₁₋₆alkyl substituted with 1-6 fluoro, and —O—C₁₋₃alkyl;and pharmaceutically acceptable salts and individual diastereomersthereof.

In the present invention it is preferred that X is selected from thegroup consisting of: —O—, —CH₂—, —S—, —SO—, and —SO₂—.

In the present invention it is more preferred that X is selected fromthe group consisting of: —O—, and —CH₂—.

In the present invention it is preferred that X is —O—.

In the present invention it is preferred that R¹ is selected from:

-   -   (1) —C₁₋₆alkyl, which is unsubstituted or substituted with 1-6        substituents where the substituents are independently selected        from: halo, hydroxy, —O—C₁₋₃alkyl, and trifluoromethyl,    -   (2) —C₀₋₆alkyl-O—C₁₋₆alkyl-, which is unsubstituted or        substituted with 1-6 substituents where the substituents are        independently selected from: halo, and trifluoromethyl,    -   (3) —C₀₋₆alkyl-S—C₁₋₆alkyl-, which is unsubstituted or        substituted with 1-6 substituents where the substituents are        independently selected from: halo, and trifluoromethyl,    -   (4) —(C₃₋₅cycloalkyl)-(C₀₋₆alkyl), which is unsubstituted or        substituted with 1-7 substituents where the substituents are        independently selected from: halo, hydroxy, —O—C₁₋₃alkyl, and        trifluoromethyl.

In the present invention it is more preferred that R¹ is C₁₋₆alkyl whichis unsubstituted or substituted with 1-5 substituents where thesubstituents are independently selected from: hydroxy, and fluoro.

In the present invention it is more preferred that R¹ is selected from:C₁₋₆alkyl, C₁₋₆alkyl-hydroxy, and C₁₋₆alkyl substituted with 1-6 fluoro.

In the present invention it is even more preferred that R¹ is selectedfrom: isopropyl, —CH(OH)CH₃, and —CH₂CF₃.

In the present invention it is preferred that R² is selected from:hydrogen, hydroxy, trifluoromethyl.

In the present invention it is more preferred that R² is selected from:hydrogen, and hydroxy.

In the present invention it is more preferred that R² is hydrogen.

In the present invention it is preferred that R³ is selected from:C₁₋₆alkyl unsubstituted or substituted with 1-6 fluoro, fluoro, chloro,bromo.

In the present invention it is more preferred that R³ is selected from:trifluromethyl, cyclopropyl, fluoro.

In the present invention it is even more preferred that R³ istrifluromethyl.

In the present invention it is preferred that R⁴ is selected from:hydrogen, and trifluoromethyl.

In the present invention it is more preferred that R⁴ is hydrogen.

In the present invention it is preferred that R⁵ is selected from:C₁₋₆alkyl unsubstituted or substituted with 1-6 fluoro, fluoro, chloro,bromo.

In the present invention it is more preferred that R⁵ is selected from:trifluromethyl, cyclopropyl, and fluoro.

In the present invention it is most preferred that R⁵ istrifluoromethyl.

In the present invention it is preferred that R⁶ is hydrogen.

In the present invention it is preferred that R⁷ is hydrogen.

In the present invention it is preferred that R⁸ is selected from:hydrogen, C₁₋₃alkyl, which is unsubstituted or substituted with 1-6fluoro, —O—C₁₋₃alkyl, fluoro, and hydroxy.

In the present invention it is more preferred that R⁸ is selected from:hydrogen, methyl, ethyl, trifluoromethyl, fluoro, and —O—CH₃.

In the present invention it is preferred that R⁹ is hydrogen.

In the present invention it is preferred that R¹⁰ is hydrogen.

In the present invention it is also preferred that R⁸ and R¹⁰ are joinedtogether by a —CH₂CH₂— chain to form a cyclopentyl ring.

In the present invention it is preferred that R²⁷ is ═O, where R²⁷ is 0and is joined via a double bond.

In the present invention it is preferred that R²⁹, R³⁰ and R³¹ are allhydrogen.

Representative compounds of the present invention include the titlecompounds of the Examples and pharmaceutically acceptable salts andindividual diastereomers thereof.

The compounds of the instant invention have at least two asymmetriccenters at the 1- and 3-positions of the cyclopentyl ring. Additionalasymmetric centers may be present depending upon the nature of thevarious substituents on the molecule. Each such asymmetric center willindependently produce two optical isomers and it is intended that all ofthe possible optical isomers and diastereomers in mixtures and as pureor partially purified compounds are included within the ambit of thisinvention. The absolute configurations of the more preferred compoundsof this invention are of the orientation where the substituents on thecyclopentyl ring are cis, i.e. as depicted:

The absolute configurations of the most preferred compounds of thisinvention are those of the orientation as depicted:

wherein the arido substituent is designated as being of the “R” absoluteconfiguration and the amine substituent is designated as being of the“S” absolute configuration (although the designation for the amidosubstituent may be specified as “R” if the priority for assignment ofthe groups at that position differs).

The independent syntheses of diastereomers and enantiomers or theirchromatographic separations may be achieved as known in the art byappropriate modification of the methodology disclosed herein. Theirabsolute stereochemistry may be determined by the x-ray crystallographyof crystalline products or crystalline intermediates which arederivatized, if necessary, with a reagent containing an asymmetriccenter of known absolute configuration.

As appreciated by those of skill in the art, halo or halogen as usedherein are intended to include chloro, fluoro, bromo and iodo.Similarly, C₁₋₈, as in C₁₋₈alkyl is defined to identify the group ashaving 1, 2, 3, 4, 5, 6, 7 or 8 carbons in a linear or branchedarrangement, such that C₁₋₈alkyl specifically includes methyl, ethyl,n-propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl, pentyl, hexyl,heptyl and octyl. Likewise, C₀, as in C₀alkyl is defined to identify thepresence of a direct covalent bond. The term “heterocycle” as usedherein is intended to include the following groups: benzoimidazolyl,benzofuranyl, benzofurazanyl, benzopyrazolyl, benzotriazolyl,benzothiophenyl, benzoxazolyl, carbazolyl, carbolinyl, cinnolinyl,furanyl, imidazolyl, indolinyl, indolyl, indolazinyl, indazolyl,isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl,naphthpyridinyl, oxadiazolyl, oxazolyl, oxetanyl, pyranyl, pyrazinyl,pyrazolyl, pyridazinyl, pyridopyridinyl, pyridazinyl, pyridyl,pyrimidyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl,tetrahydropyranyl, tetrazolyl, tetrazolopyridyl, thiadiazolyl,thiazolyl, thienyl, triazolyl, azetidinyl, 1,4-dioxanyl,hexahydroazepinyl, piperazinyl, piperidinyl, pyrrolidinyl, morpholinyl,thiomorpholinyl, dihydrobenzoimidazolyl, dihydrobenzofuranyl,dihydrobenzothiophenyl, dihydrobenzoxazolyl, dihydrofuranyl,dihydroimidazolyl, dihydroindolyl, dihydroisooxazolyl,dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl,dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl,dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl,dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl,dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl,methylenedioxybenzoyl, tetrahydrofuranyl, and tetrahydrothienyl, andN-oxides thereof.

The language “the dashed line represents a single or a double bond”refers to a dashed line used in conjunction with a solid line. Thus adashed line next to a solid line together represent either a single bondor a double bond. For instance, the depiction

refers to either

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

As used herein, “pharmaceutically acceptable salts” refer to derivativeswherein the parent compound is modified by making acid or base saltsthereof. Examples of pharmaceutically acceptable salts include, but arenot limited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. The pharmaceutically acceptable salts include theconventional non-toxic salts or the quaternary ammonium salts of theparent compound formed, for example, from non-toxic inorganic or organicacids. For example, such conventional non-toxic salts include thosederived from inorganic acids such as hydrochloric, hydrobromic,sulfuric, sulfamic, phosphoric, nitric and the like; and the saltsprepared from organic acids such as acetic, propionic, succinic,glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic,maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic,sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic,ethane disulfonic, oxalic, isethionic, and the like.

The pharmaceutically acceptable salts of the present invention can beprepared from the parent compound which contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in water or inan organic solvent, or in a mixture of the two; generally, nonaqueousmedia such as ether, ethyl acetate, ethanol, isopropanol, oracetonitrile are preferred. Suitable salts are found, e.g. inRemington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company,Easton, Pa., 1985, p. 1418.

Exemplifying the invention is the use of the compounds disclosed in theExamples and herein.

Specific compounds within the present invention include a compound whichselected from the group consisting of: the title compounds of theExamples;

and pharmaceutically acceptable salts thereof and individualdiastereomers thereof.

The subject compounds are useful in a method of modulating chemokinereceptor activity in a patient in need of such modulation comprising theadministration of an effective amount of the compound.

The present invention is directed to the use of the foregoing compoundsas modulators of chemokine receptor activity. In particular, thesecompounds are useful as modulators of the chemokine receptors, inparticular CCR-2.

The utility of the compounds in accordance with the present invention asmodulators of chemokine receptor activity may be demonstrated bymethodology known in the art, such as the assay for chemokine binding asdisclosed by Van Riper, et al., J. Exp. Med., 177, 851-856 (1993) whichmay be readily adapted for measurement of CCR-2 binding.

Receptor affinity in a CCR-2 binding assay was determined by measuringinhibition of ¹²⁵I-MCP-1 to the endogenous CCR-2 receptor on variouscell types including monocytes, THP-1 cells, or after heterologousexpression of the cloned receptor in eukaryotic cells. The cells weresuspended in binding buffer (50 mM HEPES, pH 7.2, 5 mM MgCl₂, 1 mMCaCl₂, and 0.50% BSA) with and added to test compound or DMSO and¹²⁵I-MCP-1 at room temperature for 1 h to allow binding. The cells werethen collected on GFB filters, washed with 25 mM HEPES buffer containing500 mM NaCl and cell bound ¹²⁵I-MCP-1 was quantified.

In a chemotaxis assay chemotaxis was performed using T cell depletedPBMC isolated from venous whole or leukophoresed blood and purified byFicoll-Hypaque centrifugation followed by rosetting withneuraminidase-treated sheep erythrocytes. Once isolated, the cells werewashed with HBSS containing 0.1 mg/ml BSA and suspended at 1×10⁷cells/ml. Cells were fluorescently labeled in the dark with 2 μMCalcien-AM (Molecular Probes), for 30 min at 37° C. Labeled cells werewashed twice and suspended at 5×10⁶ cells/ml in RPMI 1640 withL-glutamine (without phenol red) containing 0.1 mg/ml BSA. MCP-1(Peprotech) at 10 ng/ml diluted in same medium or medium alone wereadded to the bottom wells (27 μl). Monocytes (150,000 cells) were addedto the topside of the filter (30 PI) following a 15 min preincubationwith DMSO or with various concentrations of test compound. An equalconcentration of test compound or DMSO was added to the bottom well toprevent dilution by diffusion. Following a 60 min incubation at 37° C.,5% CO₂, the filter was removed and the topside was washed with HEBSScontaining 0.1 mg/ml BSA to remove cells that had not migrated into thefilter. Spontaneous migration (chemokinesis) was determined in theabsence of chemoattractant

In particular, the compounds of the following examples had activity inbinding to the CCR-2 receptor in the aforementioned assays, generallywith an IC₅₀ of less than about 1 μM. Such a result is indicative of theintrinsic activity of the compounds in use as modulators of chemokinereceptor activity.

Mammalian chemokine receptors provide a target for interfering with orpromoting eosinophil and/or lymphocyte function in a mammal, such as ahuman. Compounds which inhibit or promote chemokine receptor function,are particularly useful for modulating eosinophil and/or lymphocytefunction for therapeutic purposes. Accordingly, compounds which inhibitor promote chemokine receptor function would be useful in the preventionand/or treatment of a wide variety of inflammatory and immunoregulatorydisorders and diseases, allergic diseases, atopic conditions includingallergic rhinitis, dermatitis, conjunctivitis, and asthma, as well asautoimmune pathologies such as rheumatoid arthritis and atherosclerosis.

For example, an instant compound which inhibits one or more functions ofa mammalian chemokine receptor (e.g., a human chemokine receptor) may beadministered to inhibit (i.e., reduce or prevent) inflammation. As aresult, one or more inflammatory processes, such as leukocyteemigration, chemotaxis, exocytosis (e.g., of enzymes, histamine) orinflammatory mediator release, is inhibited.

In addition to primates, such as humans, a variety of other mammals canbe treated according to the method of the present invention. Forinstance, mammals including, but not limited to, cows, sheep, goats,horses, dogs, cats, guinea pigs, rats or other bovine, ovine, equine,canine, feline, rodent or murine species can be treated. However, themethod can also be practiced in other species, such as avian species(e.g., chickens).

Diseases and conditions associated with inflammation and infection canbe treated using the compounds of the present invention. In a preferredembodiment, the disease or condition is one in which the actions oflymphocytes are to be inhibited or promoted, in order to modulate theinflammatory response.

Diseases or conditions of humans or other species which can be treatedwith inhibitors of chemokine receptor function, include, but are notlimited to: inflammatory or allergic diseases and conditions, includingrespiratory allergic diseases such as asthma, particularly bronchialasthma, allergic rhinitis, hypersensitivity lung diseases,hypersensitivity pneumonitis, eosinophilic pneumonias (e.g., Loeffler'ssyndrome, chronic eosinophilic pneumonia), delayed-typehypersentitivity, interstitial lung diseases (ILD) (e.g., idiopathicpulmonary fibrosis, or ILD associated with rheumatoid arthritis,systemic lupus erythematosus, ankylosing spondylitis, systemicsclerosis, Sjogren's syndrome, polymyositis or dermatomyositis);systemic anaphylaxis or hypersensitivity responses, drug allergies(e.g., to penicillin, cephalosporins), insect sting allergies;autoimmune diseases, such as rheumatoid arthritis, psoriatic arthritis,multiple sclerosis, systemic lupus erythematosus, myasthenia gravis,juvenile onset diabetes; glomerulonephritis, autoimmune thyroiditis,Behcet's disease; graft rejection (e.g., in transplantation), includingallograft rejection or graft-versus-host disease; inflammatory boweldiseases, such as Crohn's disease and ulcerative colitis;spondyloarthropathies; scleroderma; psoriasis (including T-cell mediatedpsoriasis) and inflammatory dermatoses such an dermatitis, eczema,atopic dermatitis, allergic contact dermatitis, urticaria; vasculitis(e.g., necrotizing, cutaneous, and hypersensitivity vasculitis);eosinphilic myositis, eosinophilic fasciitis; cancers with leukocyteinfiltration of the skin or organs. Other diseases or conditions inwhich undesirable inflammatory responses are to be inhibited can betreated, including, but not limited to, reperfusion injury,atherosclerosis, certain hematologic malignancies, cytokine-inducedtoxicity (e.g., septic shock, endotoxic shock), polymyositis,dermatomyositis.

Diseases or conditions of humans or other species which can be treatedwith modulators of chemokine receptor function, include, but are notlimited to: immunosuppression, such as that in individuals withimmunodeficiency syndromes such as AIDS or other viral infections,individuals undergoing radiation therapy, chemotherapy, therapy forautoimmune disease or drug therapy (e.g., corticosteroid therapy), whichcauses immunosuppression; immunosuppression due to congenital deficiencyin receptor function or other causes; and infections diseases, such asparasitic diseases, including, but not limited to helminth infections,such as nematodes (round worms), (Trichuriasis, Enterobiasis,Ascariasis, Hookworm, Strongyloidiasis, Trichinosis, filariasis),trematodes (flukes) (Schistosomiasis, Clonorchiasis), cestodes (tapeworms) (Echinococcosis, Taeniasis saginata, Cysticercosis), visceralworms, visceral larva migraines (e.g., Toxocara), eosinophilicgastroenteritis (e.g., Anisaki sp., Phocanema sp.), and cutaneous larvamigraines (Ancylostona braziliense, Ancylostoma caninum). In addition,treatment of the aforementioned inflammatory, allergic and autoimmunediseases can also be contemplated for promoters of chemokine receptorfunction if one contemplates the delivery of sufficient compound tocause the loss of receptor expression on cells through the induction ofchemokine receptor internalization or delivery of compound in a mannerthat results in the misdirection of the migration of cells.

The compounds of the present invention are accordingly useful in theprevention and treatment of a wide variety of inflammatory andimmunoregulatory disorders and diseases, allergic conditions, atopicconditions, as well as autoimmune pathologies. In a specific embodiment,the present invention is directed to the use of the subject compoundsfor the prevention or treatment of autoimmune diseases, such asrheumatoid arthritis or psoriatic arthritis.

In another aspect, the instant invention may be used to evaluateputative specific agonists or antagonists of chemokine receptors,including CCR-2. Accordingly, the present invention is directed to theuse of these compounds in the preparation and execution of screeningassays for compounds which modulate the activity of chemokine receptors.For example, the compounds of this invention are useful for isolatingreceptor mutants, which are excellent screening tools for more potentcompounds. Furthermore, the compounds of this invention are useful inestablishing or determining the binding site of other compounds tochemokine receptors, e.g., by competitive inhibition. The compounds ofthe instant invention are also useful for the evaluation of putativespecific modulators of the chemokine receptors, including CCR-2. Asappreciated in the art, thorough evaluation of specific agonists andantagonists of the above chemokine receptors has been hampered by thelack of availability of non-peptidyl (metabolically resistant) compoundswith high binding affinity for these receptors. Thus the compounds ofthis invention are commercial products to be sold for these purposes.

The present invention is further directed to a method for themanufacture of a medicament for modulating chemokine receptor activityin humans and animals comprising combining a compound of the presentinvention with a pharmaceutical carrier or diluent.

The present invention is further directed to the use of the presentcompounds in the prevention or treatment of infection by a retrovirus,in particular, herpes virus or the human immunodeficiency virus (HIV)and the treatment of, and delaying of the onset of consequentpathological conditions such as AIDS. Treating AIDS or preventing ortreating infection by HIV is defined as including, but not limited to,treating a wide range of states of HIV infection: AIDS, ARC (AIDSrelated complex), both symptomatic and asymptomatic, and actual orpotential exposure to HIV. For example, the compounds of this inventionare useful in treating infection by HIV after suspected past exposure toHIV by, e.g., blood transfusion, organ transplant, exchange of bodyfluids, bites, accidental needle stick, or exposure to patient bloodduring surgery.

In a preferred aspect of the present invention, a subject compound maybe used in a method of inhibiting the binding of a chemokine to achemokine receptor, such as CCR-2, of a target cell, which comprisescontacting the target cell with an amount of the compound which iseffective at inhibiting the binding of the chemokine to the chemokinereceptor.

The subject treated in the methods above is a mammal, preferably a humanbeing, male or female, in whom modulation of chemokine receptor activityis desired. “Modulation” as used herein is intended to encompassantagonism, agonism, partial antagonism, inverse agonism and/or partialagonism. In a preferred aspect of the present invention, modulationrefers to antagonism of chemokine receptor activity. The term“therapeutically effective amount” means the amount of the subjectcompound that will elicit the biological or medical response of atissue, system, animal or human that is being sought by the researcher,veterinarian, medical doctor or other clinician.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients in the specified amounts, as wellas any product which results, directly or indirectly, from combinationof the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The terms “administration of” and or “administering a” compound shouldbe understood to mean providing a compound of the invention to theindividual in need of treatment.

As used herein, the term “treatment” refers both to the treatment and tothe prevention or prophylactic therapy of the aforementioned conditions.

Combined therapy to modulate chemokine receptor activity and therebyprevent and treat inflammatory and immunoregulatory disorders anddiseases, including asthma and allergic diseases, as well as autoimmunepathologies such as rheumatoid arthritis and atherosclerosis, and thosepathologies noted above is illustrated by the combination of thecompounds of this invention and other compounds which are known for suchutilities.

For example, in the treatment or prevention of inflammation, the presentcompounds may be used in conjunction with an antiinflammatory oranalgesic agent such as an opiate agonist, a lipoxygenase inhibitor,such as an inhibitor of 5-lipoxygenase, a cyclooxygenase inhibitor, suchas a cyclooxygenase-2 inhibitor, an interleukin inhibitor, such as aninterleukin-1 inhibitor, an NMDA antagonist, an inhibitor of nitricoxide or an inhibitor of the synthesis of nitric oxide, a non-steroidalantiinflammatory agent, or a cytokine-suppressing antiinflammatoryagent, for example with a compound such as acetaminophen, aspirin,codeine, embrel, fentanyl, ibuprofen, indomethacin, ketorolac, morphine,naproxen, phenacetin, piroxicam, a steroidal analgesic, sufentanyl,sunlindac, tenidap, and the like. Similarly, the instant compounds maybe administered with a pain reliever; a potentiator such as caffeine, anH2-antagonist, simethicone, aluminum or magnesium hydroxide; adecongestant such as phenylephrine, phenylpropanolamine, pseudophedrine,oxymetazoline, ephinephrine, naphazoline, xylometazoline,propylhexedrine, or levo-desoxy-ephedrine; an antiitussive such ascodeine, hydrocodone, caramiphen, carbetapentane, or dextramethorphan; adiuretic; and a sedating or non-sedating antihistamine.

Likewise, compounds of the present invention may be used in combinationwith other drugs that are used in the treatment/prevention/suppressionor amelioration of the diseases or conditions for which compounds of thepressent invention are useful. Such other drugs may be administered, bya route and in an amount commonly used therefor, contemporaneously orsequentially with a compound of the present invention. When a compoundof the present invention is used contemporaneously with one or moreother drugs, a pharmaceutical composition containing such other drugs inaddition to the compound of the present invention is preferred.Accordingly, the pharmaceutical compositions of the present inventioninclude those that also contain one or more other active ingredients, inaddition to a compound of the present invention.

Examples of other active ingredients that may be combined with acompound of the present invention, either administered separately or inthe same pharmaceutical compositions, include, but are not limited to:(a) VLA-4 antagonists such as those described in U.S. Pat. No.5,510,332, WO95/15973, WO96/01644, WO96/06108, WO96/20216, WO96/22966,WO96/31206, WO96/40781, WO97/03094, WO97/02289, WO 98/42656, WO98/53814,WO98/53817, WO98/53818, WO98/54207, and WO98/58902; (b) steroids such asbeclomethasone, methylprednisolone, betamethasone, prednisone,dexamethasone, and hydrocortisone; (c) immunosuppressants such ascyclosporin, tacrolimus, rapamycin and other FK-506 typeimmunosuppressants; (d) antihistamines (H1-histamine antagonists) suchas bromopheniramine, chlorpheniramine, dexchlorpheniramine,triprolidine, clemastine, diphenhydramine, diphenylpyraline,tripelennamine, hydroxyzine, methdilazine, promethazine, trimeprazine,azatadine, cyproheptadine, antazoline, pheniramine pyrilamine,astemizole, terfenadine, loratadine, desloratadine, cetirizine,fexofenadine, descarboethoxyloratadine, and the like; (e) non-steroidalanti-asthmatics such as β2-agonists (terbutaline, metaproterenol,fenoterol, isoetharine, albuterol, bitolterol, and pirbuterol),theophylline, cromolyn sodium, atropine, ipratropium bromide,leukotriene antagonists (zafirlukast, montelukast, pranlukast,iralukast, pobilukast, SKB-106,203), leukotriene biosynthesis inhibitors(zileuton, BAY-1005); (f) non-steroidal antiinflammatory agents (NSAIDs)such as propionic acid derivatives (alminoprofen, benoxaprofen, bucloxicacid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen,ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin,pirprofen, pranoprofen, suprofen, tiaprofenic acid, and tioxaprofen),acetic acid derivatives (indomethacin, acemetacin, alclofenac, clidanac,diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac,isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin, andzomepirac), fenamic acid derivatives (flufenamic acid, meclofenamicacid, mefenamic acid, niflumic acid and tolfenamic acid),biphenylcarboxylic acid derivatives (diflunisal and flufenisal), oxicams(isoxicam, piroxicam, sudoxicam and tenoxican), salicylates (acetylsalicylic acid, sulfasalazine) and the pyrazolones (apazone,bezpiperylon, feprazone, mofebutazone, oxyphenbutazone, phenylbutazone);(g) cyclooxygenase-2 (COX-2) inhibitors; (h) inhibitors ofphosphodiesterase type IV (PDE-IV); (i) other antagonists of thechemokine receptors, especially CCR-1, CCR-2, CCR-3, CXCR-3 and CCR-5;(j) cholesterol lowering agents such as HMG-CoA reductase inhibitors(lovastatin, sinvastatin and pravastatin, fluvastatin, atorvastatin,rosuvastatin, and other statins), sequestrants (cholestyramine andcolestipol), cholesterol absorption inhibitors (ezetimibe), nicotinicacid, fenofibric acid derivatives (gemfibrozil, clofibrat, fenofibrateand benzafibrate), and probucol; (k) anti-diabetic agents such asinsulin, sulfonylureas, biguanides (metformin), α-glucosidase inhibitors(acarbose) and glitazones (troglitazone and pioglitazone); (l)preparations of interferon beta (interferon beta-1α, interferonbeta-1β); (m) other compounds such as 5-aminosalicylic acid and prodrugsthereof, antimetabolites such as azathioprine and 6-mercaptopurine, andcytotoxic cancer chemotherapeutic agents.

The weight ratio of the compound of the present invention to the secondactive ingredient may be varied and will depend upon the effective doseof each ingredient. Generally, an effective dose of each will be used.Thus, for example, when a compound of the present invention is combinedwith an NSAID the weight ratio of the compound of the present inventionto the NSAID will generally range from about 1000:1 to about 1:1000,preferably about 200:1 to about 1:200. Combinations of a compound of thepresent invention and other active ingredients will generally also bewithin the aforementioned range, but in each case, an effective dose ofeach active ingredient should be used.

In such combinations the compound of the present invention and otheractive agents may be administered separately or in conjunction. Inaddition, the administration of one element may be prior to, concurrentto, or subsequent to the administration of other agent(s).

The compounds of the present invention may be administered by oral,parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV,intracisternal injection or infusion, subcutaneous injection, orimplant), by inhalation spray, nasal, vaginal, rectal, sublingual, ortopical routes of administration and may be formulated, alone ortogether, in suitable dosage unit formulations containing conventionalnon-toxic pharmaceutically acceptable carriers, adjuvants and vehiclesappropriate for each route of administration. In addition to thetreatment of warm-blooded animals such as mice, rats, horses, cattle,sheep, dogs, cats, monkeys, etc., the compounds of the invention areeffective for use in humans.

The pharmaceutical compositions for the administration of the compoundsof this invention may conveniently be presented in dosage unit form andmay be prepared by any of the methods well known in the art of pharmacy.All methods include the step of bringing the active ingredient intoassociation with the carrier which constitutes one or more accessoryingredients. In general, the pharmaceutical compositions are prepared byuniformly and intimately bringing the active ingredient into associationwith a liquid carrier or a finely divided solid carrier or both, andthen, if necessary, shaping the product into the desired formulation. Inthe pharmaceutical composition the active object compound is included inan amount sufficient to produce the desired effect upon the process orcondition of diseases. As used herein, the term “composition” isintended to encompass a product comprising the specified ingredients inthe specified amounts, as well as any product which results, directly orindirectly, from combination of the specified ingredients in thespecified amounts.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions, hard or soft capsules, or syrups or elixirs. Compositionsintended for oral use may be prepared according to any method known tothe art for the manufacture of pharmaceutical compositions and suchcompositions may contain one or more agents selected from the groupconsisting of sweetening agents, flavoring agents, coloring agents andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as calcium carbonate, sodium carbonate,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, corn starch, or alginic acid;binding agents, for example starch, gelatin or acacia, and lubricatingagents, for example magnesium stearate, stearic acid or talc. Thetablets may be uncoated or they may be coated by known techniques todelay disintegration and absorption in the gastrointestinal tract andthereby provide a sustained action over a longer period. For example, atime delay material such as glyceryl monostearate or glyceryl distearatemay be employed. They may also be coated by the techniques described inthe U.S. Pat. Nos. 4,256,108; 4,166,452; and 4,265,874 to form osmotictherapeutic tablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethylene-oxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds of the present invention may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials are cocoa butter and polyethyleneglycols.

For topical use, creams, ointments, jellies, solutions or suspensions,etc., containing the compounds of the present invention are employed.(For purposes of this application, topical application shall includemouthwashes and gargles.)

The pharmaceutical composition and method of the present invention mayfurther comprise other therapeutically active compounds as noted hereinwhich are usually applied in the treatment of the above mentionedpathological conditions.

In the treatment or prevention of conditions which require chemokinereceptor modulation an appropriate dosage level will generally be about0.01 to 500 mg per kg patient body weight per day which can beadministered in single or multiple doses. Preferably, the dosage levelwill be about 0.1 to about 250 mg/kg per day; more preferably about 0.5to about 100 mg/kg per day. A suitable dosage level may be about 0.01 to250 mg/kg per day, about 0.05 to 100 mg/kg per day, or about 0.1 to 50mg/kg per day. Within this range the dosage may be 0.05 to 0.5, 0.5 to 5or 5 to 50 mg/kg per day. For oral administration, the compositions arepreferably provided in the form of tablets containing 1.0 to 1000milligrams of the active ingredient, preferably 2.0 to 500, morepreferably 3.0 to 200, particularly 1, 5, 10, 15, 20, 25, 30, 50, 75,100, 125, 150, 175, 200, 250, 300, 400, 500, 600, 750, 800, 900, and1000 milligrams of the active ingredient for the symptomatic adjustmentof the dosage to the patient to be treated. The compounds may beadministered on a regimen of 1 to 4 times per day, preferably once ortwice per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, general health, sex, diet, mode and timeof administration, rate of excretion, drug combination, the severity ofthe particular condition, and the host undergoing therapy.

Several methods for preparing the compounds of this invention areillustrated in the following Schemes and Examples. Starting materialsare made by known procedures or as illustrated.

One of the principal routes used for preparation of compounds within thescope of the instant invention which bear a 1,1,3-trisubstitutedcyclopentane framework 1-8 is depicted in Scheme 1A.

According to this, 3-oxocyclopentanecarboxylic acid (1-1) which wassynthesized following a known procedure (Stetter, H., Kuhlman, H.,Liebigs Ann. Chm., 1979, 944) is esterified under standard conditions.In the case of R¹³ being a benzyl group the acid is reacted with benzylchloride in the presence of sodium carbonate in an appropriate solvent,e.g. dimethyl formamide. When R¹³ represents a tert-Butyl group, therespective ester 1-2 can be prepared by reacting the appropriate alcoholin this case tert-Butanol with acid 1-1 in the presence of sulfuricacid. Protection of the oxo-group in 1-2 can be achieved by a number ofways (Greene, T., Wuts, P. G. M., Protective Groups in OrganicChemistry, John Wiley & Sons, Inc., New York, N.Y. 1991). Theparticularly suitable dimethyl acetal protecting group can be introducedusing trimethyl orthoformate as a reagent in a suitable solvent such asdichloromethane and methyl alcohol in the presence of an acidiccatalyst. Alternatively, in the case of R¹³ being a methyl group, theacid 1-1 can be converted to 1-3 directly by using trimethylorthoformate and an acidic catalyst, such as para-Toluenesulfonic acid.An alkylation of esters 1-3 with an alkylating agent such as alkylchloride, bromide or iodide in the presence of an appropriate base suchas lithium diisopropylamide, produces intermediates 14. The esterprotecting group present in 14 can be removed by a number of ways,depending on the nature of the ester. Benzyl esters (R¹³=benzyl) can beeasily removed by catalytic hydrogenation, methyl esters (R¹³=methyl)can be hydrolyzed in the presence of an acid or base at ambient orelevated temperatures, whereas tert-Butyl esters (R¹³=tert-Butyl) can beeasily cleaved under acidic conditions. The amides 1-6 are then preparedby reaction of acids 1-5 with amines R²NH₂ in the presence of a suitablecoupling agent, such as 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimideor other agents described in the literature. The acetal protecting groupis in general removed under acidic conditions and the final chemokinereceptor modulators 1-8 can be then prepared by reaction of ketones 1-7with appropriate amines R³R⁴NH in a presence of a reducing agent such assodium triacetoxyborohydride or sodium cyanoborohydride.

The enolate generated from ester 1-3 (R¹³ being a benzyl or tert-Butylgroup) in the presence of a strong base such as lithium diisopropylamidecan be reacted with aldehydes (R^(1a)CHO)Or ketones (R^(1a)R^(2a)CO) toproduce the appropriate hydroxyalkyl substituted intermediates 1-4a asindicated in Scheme 1B. Once again the ester protecting group is removedunder conditions suitable for the particular protecting group: thecleavage of the benzyl esters can be achieved hydrogenolytically, andthe acids can be converted to the final products 1-8a as described inScheme 1, or, in the case of the tert-Butyl esters, under acidicconditions. The latter induces usually cleavage of the acetal protectinggroup as well, and the keto acids 1-9 can be prepared this way in anone-pot procedure. Their conversion to the final modulators of chemokineactivity 1-8a can be achieved as described previously.

The compounds, Which can be synthesized according to the chemistrydescribed in Schemes 1A and 1B represent diastereoisomeric mixtures(Eliel, E. E., Wilen, S. H., Stereochemistry of Organic Compounds, JohnWiley & Sons, Inc., New York), and these can be separated into theircomponents by chromatography using normal phase, reverse phase or chiralcolumns, depending on the nature of the separation. The chiralchromatographic separations are particularly suitable to obtain singleisomers.

An alternative route for preparation of compounds 1-8 and 1-8a isdetailed in Schemes 2A, 2B and 2C.

According to this, the commercially available homochiral lactam 2-1 ishydrogenated and the saturated 2-2 is treated with BOC₂O in the presenceof a suitable catalyst, e.g. N,N-dimethylamino pyridine. A basecatalyzed cleavage of the amide bond in the presence of a suitablealcohol R¹³—OH provides then the respective ester 24. The BOC-protectinggroup is removed, preferably with an acid such as HCl in a aproticsolvent, such as dioxane, to yield the amine 2-5 in a form of a salt.When this amine is mixed with benzophenone imine, the respective Schiffbase 2-6 is formed, which can be obtained in pure form by simplefiltration to remove ammonium chloride.

The enolate formed from ester 2-6 with a strong base, such as LDA can bereacted with alkyl halides R¹—X, as well as aldehydes R^(1a)CHO orketones R^(1a)R^(2a)CO to obtain intermediates 2-7a, 2-7b and 2-8a, 2-8brespectively, Scheme 2B. These reactions produce a mixture of therespective cis-(2-7a and 2-8a) and trans-(2-7b and 2-8b)diastereoisomers, which can be separated by a suitable chromatography.In most cases, normal phase flash chromatography on deactivated silicagel can be applied with success.

The desired cis diastereoisomers 2-7a and 2-8a are then treated with anacid such as HCl to aid hydrolysis of the imine group and the resultingamino group is suitably protected e.g. in a form of atert-butoxycarbonyl amide (Scheme 2C). The ester group present inintermediates 2-10a is then cleaved. The applied procedure depends onthe nature of the ester: e.g. a benzyl ester can be cleaved byhydrogenolysis, an tert-Butyl ester under aprotic acidic conditions anda alkyl ester can be hydrolyzed under either acidic or basic conditions.The formed acids are then coupled with suitable amines as describedbefore and the BOC protecting group is removed with an acid. A reductivealkylation of amines 2-13a with suitable ketones R^(3a)R^(3b)CO oraldehydes R^(3a)CHO. If necessary, a second reductive alkylation tointroduce the R⁴ substituent is then performed to yield the finalmodulators of chemokine activity 1-8b.

Intermediates 2-8a can be transformed into final products 1-8a in asequence of steps (Scheme 2D) analogous to those described in Scheme 2Cexcept that a base catalyzed hydrolysis of the ester group was found notto be suitable for the desired transformation.

The chemistry described in Schemes 2A-D offers a considerable advantagein that following these transformations products 1-8 are obtained in ahomochiral form, rendering the separation step described in Scheme 1unnecessary.

The third principal route to synthesize compounds within the scope ofthe instant invention is detailed in Scheme 3A and 3B.

According to this, the commercially available ethyl aminothiazoleacetate 3-1 is treated with benzophenone imine, preferably at elevatedtemperature. The enolate, generated from ester 3-2 with a strong base,e.g. sodium hydride is then double alkylated with 1,4-dichloro-2-butenein a suitable solvent, such as dimethoxyethane preferably in thepresence of an additional co-solvent (e.g. DMPU) to suppress undesiredside-reactions. The cleavage of the Schiff base 3-3 is accomplished asdescribed previously and the amino group in 3-4 is protected bytreatment with BOC₂O in the presence of a catalytic amount of DMAP 3-5.Addition of borane to the double bond (see March, J. Advanced OrganicChemistry, 4^(th) edition, John Wiley & Sons Inc., New York, p. 702-707)is followed by a direct pyridinium chlorochromate mediated oxidation ofthe formed adduct to produce ketones 3-6 directly, in fair yield.

The ester group present in intermediates 3-6 is then removed by a basecatalyzed hydrolysis, and the acids 3-7 are coupled to amines R²NH₂ asdiscussed previously. The last step in preparation of final compounds1-8c is a reductive amination of ketones 3-8 with amines R³R⁴NH asdetailed above. Similarly to the case described in Scheme 1A and 1B thissynthetic sequence produces mixtures of diastereoisomers, and theirseparation can be accomplished using chromatography on normal-,reverse-, or chiral phases.

The previously described synthetic sequence can be used to obtain othermodulators of chemokine activity, which carry an aromatic orheteroaromatic ring, as well as other groups which can not be introducedby direct alkylation (e.g. cyano). The required chemical transformationsare detailed in Scheme 3C.

According to this, an alkyl arylacetate is reacted with1,4-dichloro-2-butene, the olefin is hydroborated and the adductoxidized to yield 3-11 as discussed previously. A base catalyzedhydrolysis is followed by an amide forming step, and the final amines1-8c are then prepared in an reductive amination step analogous to that,described above. Similarly to chemistry depicted in Schemes 3A and 3B,the sequence described in Scheme 3C yields a mixture ofdiastereoisomers. These are then separated into single isomers by meansdescribed previously.

In the cases discussed so far, the formation of the amide bond alwayspreceded the reductive amination step. In some instances, however, itwas advantageous to reverse this order (Scheme 4).

According to this, the amine intermediate 2-9a is reductively alkylatedwith an appropriate ketone R^(3a)R^(3b)CO and the resultant secondaryamine is protected e.g., as a trifluroacetamide. The ester group is nowcleaved (hydrogenolysis if R¹³ is a benzyl group) and the amide isattached as described above. A reductive or base catalyzed removal ofthe trifluoroacetyl protecting group then affords the desired modulatorsof chemokine activity 1-8e.

The amines 5-1 incorporated into the amide portion of 1-8 often containa substituted benzyl group (Scheme 5). Some of these, e.g. the3,5-bistrifluoromethylbenzylamine (R⁵═CF₃),3-fluoro-5-trifluoromethylbenzylamine (R⁵═F), or others can be obtainedcommercially, others are available through synthesis, Schemes 6, 7.

An example of such a synthesis is depicted in Scheme 6. According tothis, the commercially available 3-trifluoromethyl-5-amino bromobenzene(6-1) is converted to the corresponding nitrile using zinc cyanide inthe presence of palladium, and a Sandmeyer reaction is then used toproduce the respective halide 6-3, R⁵═Cl, I. The reduction of thenitrile in the presence of an aromatic halide to the corresponding aminecan be successfully accomplished e.g. with borane in TMF.

Additional examples of benzyl amines incorporated into the amide portionof compounds within the scope of the instant invention, as well as theirsyntheses are further described in the Experimental section.

The amine portion of the target compounds 1-8 can be, as detailed above,prepared either from reductive amination of the corresponding ketones1-7 and 1-7a with appropriate amines 7-1, or by a reductive alkylationof amines 2-9a and 2-13a with the corresponding ketones 7-3, Scheme 7.The amines 7-1 as well as ketones 7-3 typically represent alicyclicstructures, some of which, e.g. tetrahydropyran-4-one and a number oflower both cyclic and alicyclic ketones, can be obtained commercially,others have to be synthesized either by known or newly developedprocedures.

A synthesis of some of the ketones 7-3 is detailed in Scheme 8.According to this, a commercially available5,6-dihydro-4-methoxy-2H-pyran (8-1) is treated with m-chloroperbenzoicacid to affect epoxidation of the double bond. This in turn reacts withthe formed chlorobenzoic acid and the epoxide ring opening furnishes theketone 8-2. Its protection in the form of an acetal is followed byremoval of the ester. An alkylation of the secondary alcohol with anappropriate alkyl halide R⁶X in a presence of a base affords the ether8-5. Deprotection of the acetal under acidic conditions affords thedesired ketones 8-6. In this manner, besides the3-hydroxy-tetrahydropyran-4-one, a number of 3-alkoxyderivatives can besynthesized. Further details, as well as examples are described in theExperimental section.

An example of preparation of amines 7-1, in this case3-methyl-4-amino-tetrahydropyrane is detailed in Scheme 9. According tothis, 3-methyltetrahydropyran-4-one, which can be synthesized fromtetrahydropyran-4-one following a previously published procedure (J. Am.Chem. Soc., 1991, 113, 2079-2089) is reacted with benzhydrylamine underreductive amination conditions.

This transformation yields the two respective diastereoisomeric pairswith trans-(9-3a) and cis (9-3b) relative stereochemistry in a ratio ofapproximately 1 to 9. The major cis-racemic pair (9-3b) is subjected tohydrogenolysis to aid removal of the benzhydryl group (Greene, T., Wuts,P. G. M., Protective Groups in Organic Chemistry, John Wiley & Sons,Inc., New York, N.Y. 1991) and the free amine 9-4 is then reacted withbenzyloxycarbonyl chloride in the presence of a suitable base, e.g.diisopropylethylamine. The two enantiomers contained within the racemate9-5 can be then separated using chiral preparative chromatography. ADAICEL® Chiralpak AD polysacharide type preparative column (ChiralTechnologies, 730 Springdale Drive, P.O. Box 564, Exton, Pa. 19341) canbe used for the desired separation with great success. The fastereluting enantiomer 9-6b was shown to have the absolute stereochemistry(3R,4S) by derivatization and NMR analysis as well as single crystalX-Ray diffraction analysis (Eliel, E. E., Wilen, S. H., Stereochemistryof Organic Compounds, John Wiley & Sons, Inc., New York). The CBZprotecting group is then removed hydrogenolytically to yield 9-7 inexcellent yield.

Additional details as well as examples of preparation of amines 7-1 canbe found in the Experimental section.

In some cases the order of carrying out the foregoing reaction schemesmay be varied to facilitate the reaction or to avoid unwanted reactionproducts. The following examples are provided for the purpose of furtherillustration only and are not intended to be limitations on thedisclosed invention.

Concentration of solutions was generally carried out on a rotaryevaporator under reduced pressure. Flash chromatography was carried outon silica gel (230-400 mesh). MPLC refers to medium pressure liquidchromatography and was carried out on a silica gel stationary phaseunless otherwise noted. NMR spectra were obtained in CDCl₃ solutionunless otherwise noted. Coupling constants (J) are in hertz (Hz).Abbreviations: diethyl ether (ether), triethylamine (TEA),N,N-diisopropylethylamine (DIEA) saturated aqueous (sat'd), roomtemperature (rt), hour(s) (h), minute(s) (min).

Intermediate 1

Step A

A solution of 3-oxo-cyclopentanecarboxylic acid (20 g, 160 mmol,Stetter, H., Kuhlman, H., Liebigs Ann. Chemie, 1979, 7, 944-9) in MeOH(200 mL) and trimethyl orthoformate (85 mL, 780 mmol) followed by TsOH(3.0 g, 16 mmol) was added. The reaction mixture was stirred at roomtemperature for 4 h and concentrated in vacuo. The residue was dilutedwith ether, washed with saturated NaHCO₃, brine, and dried overanhydrous MgSO₄. The crude product was purified by flash chromatography(ether:pentane/25:75,) to yield the desired product (21.52 g, 73%). ¹HNMR (500 MHz, CDCl₃): 3.68 (s, 3H), 3.21 (d, J=9.9 Hz, 6H), 2.89 (p,J=8.5 Hz, 1H), 2.14-2.05 (m, 2H), 2.02-1.80 (m, 4H).Step B

A flame-dried 500 mL round-bottomed flask, was charged with dry THF (150mL) and diisopropylamine (19.2 mL, 137 mmol) was added. The solution wascooled to −78° C. and n-butyllithium (55 mL, 140 mmol, 2.5 M solution inhexanes) was added via syringe, followed by neat ester from the previousstep (21.52 g, 114.4 mmol). The reaction mixture was stirred at −78° C.for 30 minutes and 2-iodopropane (34.3 mL, 343.2 mmol) was added. Afterthe reaction was stirred for another 20 minutes at −78° C., it was keptin a refrigerator (+5° C.) overnight. The reaction was quenched with 10%citric acid and extracted with ether (3×). The combined organic layerswere washed with H₂O, brine, dried over anhydrous MgSO₄, andconcentrated. The residue was purified by flash chromatography(ether:pentane/20:80) to yield the desired product (16.74 g, 63.6%). ¹HNMR (400 MHz, CDCl₃): 3.69 (s, 3H), 3.18 (d, J=20.5 Hz, 6H), 2.57 (d,J=13.9 Hz, 1H), 2.29 (m, 1H), 1.90 (p, J=6.8 Hz, 1H), 1.81 (m, 2H), 1.65(m, 2H), 0.89 (q, J=11.9 Hz, 6.8 Hz, 6H).Step C

The ester from the previous step (16.7 g, 72.7 mmol) was dissolved inethanol (30 mL) and a solution of NaOH (11 g, 280 mmol) in H₂O (30 mL)was added. The reaction mixture was refluxed for 3 days, cooled to roomtemperature and acidified with concentrated HCl. The alcohol wasevaporated under reduced pressure and the residue was extracted withdichloromethane (5×). The combined organic layers were dried overanhydrous MgSO₄ and concentrated in vacuo to yield 11.07 g (89%) of thedesired acid Intermediate 1. ¹H NMR (500 MHz, CDCl₃): 2.70 (d, J=18.1Hz, 1H), 2.44-2.39 (m, 1H), 2.30-2.15 (m, 2H), 2.14 (dd, J=18.1 Hz, 1.0Hz, 1H), 2.06 (p, J=6.9 Hz, 1H), 1.98 (m, 1H), 0.98 (dd, J=11.4 Hz, 6.9Hz, 6H).

Intermediate 2

Step A

To a solution of tetrahydro-4H-pyran-4-one (5 g, 50 mmol) anddiphenylmethylamine (8.4 mL, 50 mmol) in DCM (250 mL) was added 4 Åpowdered molecular sieves followed by NaB(OAc)₃H (32 g, 150 mmol). Thereaction mixture was stirred at room temperature overnight. It wasfiltered through celite, the filtrate was washed with saturated NaHCO₃(4×), dried over MgSO₄, and concentrated in vacuo to yield a crudeproduct (13.25 g, 99.9%). ¹H NMR (400 MHz, CDCl₃): 7.42 (bd, J=7.0 Hz,4H), 7.32 (bt, J=7.2 Hz, 4H), 7.24 (bt, J=7.3 Hz, 2H), 5.07 (s, 1H),3.96 (dt, J=11.1 Hz, 3.5 Hz, 2H), 3.33 (td, J=11.5 Hz, 2.1 Hz, 2H), 2.66(m, 1H), 1.93 (m, 2H), 1.54 (bs, 1H), 1.44 (m, 2H).Step B

A mixture of the amine from previous step (13.2 g, 49.4 mmol), 4NHCl/dioxane (12.5 mL, 49.4 mmol), Pd/C 10% (1.1 g), dioxane (30 mL), andethanol (120 mL) was hydrogenated at 35 psi pressure overnight on a Parrapparatus. The catalyst was filtered off and the filtrate wasconcentrated to dryness. The residue was triturated with DCM, and theprecipitate was filtered, and dried to yield 4.91 g (72%) ofIntermediate 2 in a form of a hydrochloride salt. ¹H NMR (400 MHz,CD₃OD): 3.99 (dd, J=12.1 Hz, 5.1 Hz, 2H), 1.89 (td, J=11.9 Hz, 2.1 Hz,2H), 3.38-3.32 (m, 1H), 1.96-1.92 (m, 2H), 1.70-1.59 (m, 2H).

Intermediate 3

A flame-dried 250 mL round-bottomed flask was charged with5,6-dihydro-4-methoxy-2H-pyran (5.00 g, 43.8 mmol) and dry DCM (150 mL).Na₂HPO₄ (22.4 g, 158 mmol) was added and the mixture was cooled to 0° C.in an ice-bath. A solution of m-CPBA (13.6 g, 78.8 mmol) in DCM (30 mL)was added slowly via syringe. The reaction mixture was then allowed towarm up to room temperature. Upon completion of reaction, the mixturewas diluted with DCM, washed with H₂O (3×) and saturated NaCl, driedover Na₂SO₄, and concentrated in vacuo to yield the desired Intermediate3 (8.36 g, 75.0%). ¹H NMR (500 MHz, CDCl₃): 8.06 (t, J=1.7 Hz, 1H), 7.97(dt, J=0.8 Hz, 1.4 Hz, 1H), 7.58 (dq, J=8.0 Hz, 1.1 Hz, 1H), 7.42 (t,J=7.8 Hz, 1H), 5.53 (ddd, J=10.6 Hz, 7.1 Hz, 1.0 Hz, 1H), 4.48 (ddd,J=10.8 Hz, 6.9 Hz, 1.4 Hz, 1H), 4.34 (m, 1H), 3.74 (m, 2H), 2.67 (m,1H), 2.62 (dt, J=14.4 Hz, 1.8 Hz, 1H).

Intermediate 4

Step A

Intermediate 3 (200 mg, 0.787 mmol) was dissolved in DCM (5 mL) and MeOH(5 mL) before trimethyl orthoformate (8.68 mL, 7.87 mmol) and TsOH (15mg, 0.0787 mmol) were added. The reaction mixture was stirred at roomtemperature for two days, concentrated to dryness and purified bypreparative TLC to yield the desired acetal (161 mg, 68.2%). ¹H NMR (500MHz, CDCl₃): 8.07 (t, J=1.7 Hz, 1H), 7.99 (dt, J=7.8 Hz, 1.3 Hz, 1H),7.56 (dq, J=8.0 Hz, 1.1 Hz, 1H), 7.41 (t, J=8.0 Hz, 1H), 7.12 (d, J=1.9Hz, H), 4.00 (dd, J=12.8 Hz, 1.8 Hz, 1H), 3.93 (dd, J=11.5 Hz, 3.9 Hz,1H), 3.83 (dd, J=12.9 Hz, 1.4 Hz, 1H), 3.58 (m, 1H), 3.29 (s, 3H), 3.19(s, 1H), 2.13 (m, 1H), 1.95 (dd, J=14.1 Hz, 2.0 Hz, 1H).Step B

The ester from previous step (160 mg, 0.533 mmol) was dissolved in MeOH(2 mL) and a solution of NaOMe (0.5 M in MeOH, 1.3 mL) was added. Thereaction was stirred for 1.5 h. The mixture was concentrated in vacuoand the crude product was used in the next step without furtherpurificationStep C

NaH (50 mg, 1.1 mmol) was suspended in THF (5 mL), and the crude alcoholfrom previous step (90 mg, 0.53 mmol) was added, followed by allylbromide (461 μL, 5.33 mmol). The reaction mixture was stirred at roomtemperature overnight, and concentrated to dryness. The residue wasdiluted with ether, washed with saturated NaCl, dried over MgSO₄, andconcentrated in vacuo. The crude product was purified by preparative TLCto yield 81 mg, (75% for last two steps) of the desired product. ¹H NMR(400 MHz, CDCl₃): 5.97 (m, 1H), 5.32 (dq, J=17.2 Hz, 1.6 Hz, 1H), 5.21(dd, J=10.1 Hz, 1.2 Hz, 1H), 4.21 (m, 1H), 4.08 (m, 1H), 3.94 (dd, 12.1Hz, 2.9 Hz, 1H), 3.80 (td, J=11.1 Hz, 3.9 Hz, 1H), 3.61 (dd, J=12.3 Hz,1.6 Hz, 1H), 3.51 (dt, 1=11.5 Hz, 2.7 Hz, 1H), 3.39 (m, 1H), 3.26 (s,3H), 3.24 (s, 3H), 2.02 (m, 1H), 1.75 (md, J=14.0 Hz, 1H).Step D

The previously described acetal (80 mg, 0.40 mmol) was dissolved in a10% solution of TFA in DCM and stirred at room temperature for 1.5 h.The solvent was removed in vacuo and the concentrate was diluted withether and washed with saturated NaHCO₃ (5×). The combined aqueous layerwas back extracted with ether (1×). The combined organic layers weredried over MgSO₄ and concentrated in vacuo to yield Intermediate 4 (61mg, 98%). ¹H NMR (400 MHz, CDCl₃) δ 5.92 (m, 1H), 5.27 (m, 2H), 4.29(tq, J=0.5 Hz, 1.6 Hz, 1H), 4.23 (dq, J=6.3 Hz, 1.6 Hz, 1H), 4.14 (m,1H), 4.05 (tq, J=6.1 Hz, 1.4 Hz, 1H), 3.99 (m, 1H), 3.73 (m, 1H), 3.58(app. q, 1H), 3.61 (m, 2H).

Intermediate 5

Intermediate 5 was prepared according to the procedure described in J.Am. Chem. Soc., 1991, 113, 2079-2089.

Intermediate 6

Step A

DMF (10 mL) was deoxygenated (nitrogen purge, 30 minutes) and3-amino-5-bromobenzotrifluoride (500 mg, 2.08 mmol) in DMF was added.The solution was purged with nitrogen for another 10 minutes and zinccyanide (147 mg, 1.25 mmol) was added followed bytetrakis(triphenylphospine)palladium (96 mg, 0.083 mmol). Nitrogen waspassed through for another 15 minutes then it was heated to 80° C. in asealed tube overnight. The reaction mixture was diluted with ethylacetate, washed with an ammonium hydroxide solution (2×), andconcentrated in vacuo. The crude product was purified by preparative LC(ethyl acetate:hexanes/30:70) to yield 289 mg (74%) of the desiredproduct ¹H NMR (400 MHz, CDCl₃): 7.25 (d, J=0.6 Hz, 1H), 7.08 (s, 1H),7.06 (d, J=0.9 Hz, 1H). L-C-MS for C₁₀H₈F₃N₃ [M+CH₃CN]⁺ calculated227.07, found 227.8.Step B

A mixture of Copper(II) chloride (250 mg, 1.34 mmol), tert-butylnitrite(217 μL, 1.61 mmol), and anhydrous acetonitrile (7 mL), was cooled to 0°C. and the nitrile from previous step (415 mg, 1.55 mmol) in anhydrousacetonitrile (2 mL) was slowly added. The reaction mixture was heated to65° C. and monitored by TLC. When the reaction was complete, the mixturewas cooled to room temperature, poured into 20% aqueous HCl andextracted with ether. The organic layer was washed with 20% aqueous HCl,dried over anhydrous MgSO₄, and concentrated in vacuo. The crude productwas used in next step without further purification.Step C

The nitrile from previous step (270 mg, 1.34 mmol) was dissolved in THF(1 mL) and 1M Borane in THF (6.70 mL, 6.70 mmol) was added. The mixturewas stirred at room temperature overnight and concentrated in vacuo. Theresidue was picked up in a solution of 1% HCl (4N in dioxane) inmethanol and heated at 50° C. overnight. Solvent was stripped off andthe residue was dissolved in 1% HCl in methanol. This process wasrepeated three times to yield crude Intermediate 6 (212 mg, 64.4%). ¹HNMR (400 MHz, CDCl₃): 7.80 (m, 3H), 4.22 (s, 2H).

Intermediate 7

Step A

3-Amino-5-trifluoromethylbenzonitrile (500 mg, 2.69 mmol) was added to amixture of concentrated HCl (5 mL) and water (5 mL) (solution A). Sodiumnitrite (342 mg, 4.95 mmol) was dissolved in water (5 mL) (solution B).The two solutions were cooled to 0° C. separately before solution B wasadded slowly to solution A. At end of addition, KI-starch paper was usedto test presence of nitrous acid. A solution of KI (765 mg, 4.61 mmol)in water (5 mL) was added and stirred for 30 minutes followed by heatingto 100° C. until no nitrogen gas evolution was seen. The mixture wasallowed cool to room temperature and extracted with ether (2×), driedover anhydrous NaSO₄, and concentrated in vacuo. Crude product waspurified by preparative TLC (ethyl acetate:hexanes/40:60,) to yield 580mg (72%) of the desired product. ¹H NMR (300 MHz, CDCl₃): 8.18 (d,J=1.38 Hz, 2H), 7.89 (t, J=0.73 Hz, 1H).Step B

Intermediate 7 was prepared as detailed in Intermediate 6, Step 3. ¹HNMR (400 MHz, CD₃OD): 8.16 (s, 1H), 8.09 (s, 1H), 7.83 (s, 1H), 4.19 (s,2H).

Intermediate 8

Intermediate 1 (2.5 g, 15 mmol), bis(trifluoromethyl)benzylamine (4.11g, 14.7 mmol), DIEA (3.8 mL, 22 mmol), 1-hydroxy-7-azabenzotriazole (2.0g, 15 mmol) and EDC (4.23 g, 22.0 mmol) were dissolved in DCM (100 mL)and stirred at room temperature overnight. The reaction mixture waswashed with 1N HCl (2×), saturated NaHCO₃, H₂O (2×) and brine (1×). Itwas dried over anhydrous MgSO₄, and concentrated in vacuo. The crudeproduct was purified by flash chromatography (EtOAc:Hexanes/40:60) toyield Intermediate 8 (3.87 g, 66.6%). ¹H NMR (500 MHz, CDCl₃): 7.81 (s,1H), 7.74 (s, 2H), 6.16 (bs, 2H), 2.78 (bd, J=18.07 Hz, 1H), 2.40 to2.20 (bm, 4H), 2.08 to 1.98 (m, 2H), 0.99 (d, J=6.86 Hz, 3H), 0.97 (d,J=6.87 Hz, 3H).

Intermediate 9

Step A

50 g (0.46 mol) of (1S,4R)-(+)-2-azabicyclo[2.2.1]hept-5-en-3-one in 200mL of methanol containing 2.5 g of Pd/C (10%) was hydrogenated on a Parrshaker under 50 psi of hydrogen for 1 h. The catalyst was removed byfiltration through a pad of celite. The filtrates were evaporated andthe residue was dried in vacuum. The resulting white solid (50 g) wasdissolved in 200 mL of methylene chloride and 110 g (0.50 mol) ofdi-tert-butyl dicarbonate and 1.0 g of DMAP were added. The reactionmixture was stirred at room temperature overnight and then loaded on asilica gel column, eluted with 10% EtOAc/Hexane. The title compound (83g, 86%) was obtained as a white solid. ¹NMR (400 MHz, CDCl₃): 1.40 (d,1H), 1.51 (s, 9H), 1.70-1.95 (m, 5H), 2.84 (m, 1H), 4.50 (m, 1H).Step B

To a stirred mixture of 63.0 g (0.30 mol) of(1S,4R)-(+)-N-BOC-2-azabicyclo[2.2.1]hept-3-one and 32 g (0.30 mol) ofbenzyl alcohol in 200 mL of THF under nitrogen was added 2.8 g (0.30mol) of lithium hydride in multiple portions. The resulting mixture wasstirred overnight. TLC showed a complete conversion. The entire mixturewas poured into a stirred mixture of ice-water/EtOAc (500 mL). Theorganic phase was separated and washed with water (2×200 mL), dried overNa₂SO₄, evaporated and Dried in vacuum. The title compound (95.5 g,100%) was obtained as a white solid. ¹H NMR (400 MHz, CDCl₃): 1.44 (s,9H), 1.60 (m, 1H), 1.72 (m, 1H), 1.95 (m, 3H), 2.24 (m, 1H), 2.90 (m,1H), 4.08 (m, 1H), 4.98 (broad, 1H), 5.13 (s, 2H), 7.38 (m, 5H).Step C

A mixture of 96 g (0.30 mol) of(1S,3R)-benzyl-(N-BOC-3-amino)-cyclopentanecarboxylate and 300 mL of 4NHCl in dioxane was stirred for 1 h. The solvent was removed underreduced pressure, and the residue was dried under high vacuum overnightand then suspended in 300 mL of CH₂Cl₂. To this suspension was added54.4 g of benzophenone imine. The resulting mixture was stirredovernight. The precipitate was removed by filtration and the filtrateswere washed with brine, dried over Na₂SO₄, evaporated and dried invacuum.

The title compound was obtained as a light yellow oil (116.0 g, 100%).¹H NMR (400 MHz, CDCl₃): 1.80 (m, 1H), 1.95 (m, 2H), 2.15 (m, 2H), 2.50(m, 1H), 2.89 (m, 1H), 3.61 (m, 1H), 5.20 (s, 2H), 7.18 (d, 2H), 7.38(m, 8H), 7.47 (m, 3H), 7.64 (d, 2H).Step D

To a flame-dried 500 mL round-bottomed flask, was added dry THF (130mL). The solvent was cooled to −78° C. before diisopropylamine (10.5 mL,75.2 mmol), 2.5 M n-butyllithium (30 mL, 75 mmol), and a solution of theproduct prepared in Step C (25 g, 65 mmol) in THF (20 mL), were addedsequentially. The reaction mixture was stirred at −78° C. for 30 minutesbefore acetone (14.4 mL, 196 mmol) was added. After the reaction wasstirred for another h, the mixture was quenched with saturated NH₄Cl,extracted with ether, dried over MgSO₄, and concentrated. The crudeproduct was purified by MPLC (EtOAc:Hexanes/25:75). Cis and transisomers were resolved with cis being the desired isomer (cis, 6.8 g;trans, 3.47 g). %). Cis isomer: ¹H NMR (400 MHz, CDCl₃): 7.58 (m, 2H),7.48-7.28 (m, 11H), 7.14 (m, 2H), 5.22 (s, 2H), 3.78 (p, J=12.1 Hz, 6.2Hz, 1H), 3.46 (s, 1H), 2.56-2.50 (m, 1H), 2.27 (dd, J=13.9 Hz, 5.9 Hz,1H), 2.08 (dd, J=13.8 Hz, 6.6 Hz, 1H), 1.92 (m, 1H), 1.83-1.69 (m, 2H),1.09 (d, J=14.0 Hz, 6H).Step E

The imine from previous step (6.8 g, 15 mmol) was dissolved in THF (50mL) before 2 N aqueous HCl (50 mL) was added. The reaction mixture wasstirred and monitored by TLC. After completion of reaction, the mixturewas concentrated in vacuo to remove THF. The aqueous layer wasbasisified to pH 9.0 with saturated Na₂CO₃ solution and extracted withDCM. The organic layer was dried over MgSO₄ and di-tert-butyldicarbonate (4.4 g, 20 mmol) was added. The reaction was stirred at roomtemperature overnight before being extracted with DCM, dried over MgSO₄,and concentrated in vacuo. The crude product was purified by columnchromatography to yield (2.9 g, 50%). ¹H NMR (400 MHz, CDCl₃) 7.39 (m,5H), 5.20 (s, 2H), 4.62 (bs, 1H), 4.13 (b, 1H), 3.40 (s, 1H), 2.25 (dd,J=14.5 Hz, 8.1 Hz, 1H), 2.16 (m, 1H), 2.01 (m, 2H), 1.89 (m, 1H), 1.44(s, 9H), 1.18 (s, 6H).Step F

A mixture of the benzyl ester form previous step (2.9 g), Pd/C (300 mg),and ethanol (50 mL) were placed on a Parr-shaker under 50 psi pressureovernight. The mixture was filtered through celite and concentrated invacuo to yield the desired product (2.01 g, 91.0%). ¹H NMR (500 MHz,CDCl₃): 6.56 (s, ½H), 5.17 (s, ½H), 4.00 (d, J=43.3 Hz, 1H), 2.40-1.70(m, 6H), 1.46 (b, 9H), 1.27 (b, 6H).

Intermediate 10

Step A

A mixture of (1S)-(+)-2-azabicyclo[2.2.1]hept-5-en-3-one (10.3 g, 94.4mmol) in EtOAc (200 mL) and 10% Pd/C (0.5 gm), was hydrogenated at roomtemperature under a hydrogen balloon. After 24 h the reaction mixturewas filtered and evaporated leaving behind 10.4 g (100%) of a productthat was taken in 250 mL methanol and HCl (12M, 6 mL). The resultantmixture was stirred at RT, until the reaction was complete (72 h).Evaporation of methanol followed by drying under high vacuum, yieldedthe title compound as an off white solid (16.0 g, 96%).

¹H NMR (D₂O, 500 MHz): 3.70 (s, 3H), 3.01 (m, 1H), 2.38 (m, 1H),2.16-1.73 (m, 6H).Step B

To a suspension of the intermediate from step A (10.2 g, 56.8 mmol) indry dichloromethane (200 mL) was added benzophenone imine (10.2 g, 56.8mmol) at room temperature and the resultant mixture was stirred for 24h. The reaction mixture was filtered and the filtrate was evaporated, toleave behind a yellow oil that was triturated with ether (100 mL),filtered and evaporated. This operation was repeated twice to ensurethat the product was free of ammonium chloride impurities. The resultantoil was thoroughly dried under vacuum to yield the title compound (18.03g, >100%) and required no further purification. ¹H NMR (CDCl₃, 500 MHz):7.5-7.18 (m, 10H), 3.75 (m, 1H), 3.7 (s, 3H), 2.78 (m, 1H), 2.26-1.71(m, 6H).Step C

To a solution of LDA (prepared from diisopropylamine (7.7 g, 76.1 mmol)and n-butyllithium (30.4 mL, 2.5 M in hexane, 76 mmol) in THF (120 mL)at −78° C. was added the ester from Step B (18.0 g, 58.6 mmol). Theresultant burgundy colored solution was stirred for 20 min. after whichit was quenched with 2-iodopropane (14.9 gm, 88.0 mmol). The reactionmixture was gradually warmed over 3 h to 0° C. and this temperature wasmaintained for an additional 3 h. Reaction was quenched with water andextracted with EtOAc. The organic layer was washed with water, brine,dried (anhydrous magnesium sulfate) and concentrated to yield an oil. Tothe solution of the crude Schiff base (20.0 g) in THF (100 mL) was addedHCl (5.0 mL, 12 M) and was allowed to stir at room temperature for 3 h.After the removal of all volatiles, the hydrochloride salt was taken upinto dichloromethane (250 mL), and a saturated solution of sodiumbicarbonate (250 mL) and di-tert-butyl dicarbonate (26.0 g, 1.4 Eq.)were added. The resultant mixture was vigorously stirred overnight atRT. The organic layer was separated and washed with water, brine, dried(anhydrous magnesium sulfate) and concentrated to yield an oil.Purification by flash column chromatography (eluent:hexane:EtOAc/19:1)gave the desired product (4.91 g, 30%). ¹H NMR (500 MHz, CDCl₃): 4.79(br, 1H), 4.01 (m, 1H), 3.71 (s, 3H), 2.18-1.60 (m, 6H), 1.44 (s, 9H),0.87 (d, J=6.9 Hz, 3H), 0.86 (d, J=6.9 Hz, 3H).Step D

To a solution of the ester from the previous step (4.91 g, 17.2 mmol) inMeOH (100 mL) was added a solution of LiOH (3.6 g, 85 mmol) in water (20mL) and THF (10 mL). The resultant mixture was heated at 80° C. untilthe reaction was complete (18 h). Methanol was removed in vacuo and thecrude product was taken up with water/EtOAc (200 mL, 1:4) and cooled to0° C. The acidity of the mixture was adjusted to pH 6. The EtOAc layerwas separated, washed with water, brine, dried (anhydrous magnesiumsulfate) and concentrated to yield an oil. Purification by flash columnchromatography (eluent:hexane:EtOAc/1:1+2% AcOH) gave Intermediate 10(3.9 g, 84%). ¹H NMR (500 MHz, CDCl₃): 11.36 (br, 1H), 6.49 (br, 1H),4.83 (m, 1H), 3.71 (s, 3H), 2.30-1.55 (m, 6H), 1.46 (s, 9H), 0.94 (d,J=6.9 Hz, 3H), 0.933 (d, J=6.9 Hz, 3H).

Intermediate 11

Step AProcedure A

A solution of 3-oxo-cyclopentane carboxylic acid (Stetter, H., Kuhlmann,H. Liebigs Ann. Chem., 1979, 7, 944-9) (5.72 g, 44.6 mmol) indichloromethane (30 mL) was treated withN,N′-di-iso-propyl-O-tert-Butyl-iso-urea (21.2 mL, 89.3 mmol) and thereaction mixture was stirred at ambient temperature overnight. Theprecipitated N,N′-di-iso-propyl urea was filtered off, the filtrateconcentrated in vacuo and the residue was purified by distillation(b.p.: 125-129° C. @ 18 mmHg) to yield 4.7446 g (58%) of the pureproduct. ¹H NMR (500 MHz, CDCl₃): 3.02 (p, J=7.8 Hz, 1H), 2.05-2.50 (m,6H), 1.45 (s, 9H). ¹³C NMR (125 MHz, CDCl₃): 217.00, 173.47, 80.99,41.88, 41.14, 27.94, 26.57.

Procedure B

A 2 L round RBF was charged with anhydrous magnesium sulfate (113 g, 940mmol) and dichloromethane (940 mL). While stirring, the suspension wastreated with concentrated sulfuric acid (12.5 mL, 235 mmol) followedafter 15 minutes by 3-oxo-cyclopentane carboxylic acid (30.1 g, 235mmol). After stirring for 15 minutes, tert-butanol (87 g, 1.2 mol) wasadded. The reaction vessel was closed with a stopper to aid retention ofisobutylene, and stirred at ambient temperature for 72 h The solid wasfiltered off through a plug of Celite and the volume of the filtrate wasreduced to approximately 500 mL, and washed with a saturated solution ofsodium bicarbonate (2×150 mL). The organic phase was dried withanhydrous magnesium sulfate, filtered, and the solvent was removed bydistillation at reduced pressure (180 mmHg). The crude product waspurified by distillation to yield 39.12 g (90%) of pure product.Step B

A solution of tert-Butyl 3-oxocyclopentane carboxylate (11.54 g, 62.64mmol) in dichloromethane (200 mL) was treated with trimethylorthoformate (41.4 mL, 251 mmol) in the presence of p-toluenesulfonicacid (400 mg) and stirred at room temperature for 48 h. The darkreaction mixture was poured onto a saturated solution of sodiumbicarbonate, and the crude product was extracted with dichloromethane.The combined organic extracts were dried with anhydrous magnesiumsulfate, the solvent was removed in vacuo, and the crude product waspurified by distillation (b.p.: 104° C. @ 4 mmHg) to yield 12.32 g (85%)of the desired product. ¹H NMR (500 MHz, CDCl₃): 3.21 (s, 3H), 3.20 (s,3H), 2.80 (m, 1H), 2.10 to 1.80 (bm, 6H), 1.46 (s, 9H). ¹³C NMR (125MHz, CDCl₃): 174.9, 111.2, 80.3, 67.8, 49.2, 42.5, 37.4, 33.8, 28.3,22.0.Step C

A solution of diisopropylaamine (5.6 mL, 40 mmol) in dry tetrahydrofuran(40 mL) was cooled to −78° C. and it was treated with n-butyllithium (16mL, 40 mmol, 2.5M solution in hexanes). The neat ester from the previousstep (5.8 g, 25 mmol) was added via syringe, and the enolate was allowedto form for 30 minutes at −15° C. The temperature of the reactionmixture was lowered to −78° C. once again, and acetone (5.5 mL, 75 mmol)was added via syringe. The reaction was allowed to proceed at −15° C.overnight, and it was quenched by pouring the mixture onto 150 mL of 10%aqueous citric acid. The crude product was extracted into diethyl ether,the combined extracts were dried and the solvent was removed in vacuo.The crude product (8.31 g) was further purified by column chromatography(Silica gel, ethyl acetate+hexanes/1:1) to yield 4.31 g (60%) of pureproduct. ¹H NMR (500 MHz, CDCl₃): 3.21 (s, 3H), 3.18 (s, 3H), 2.46 (d,J=14.2 Hz, 1H), 2.20 (m, 1H), 1.99 (d, J=13.96 Hz), 1.85 (m, 3H), 1.50(s, 9H), 1.21 (bs, 6H). ¹³C NMR (125 MHz, CDCl₃): 175.9, 110.4, 81.8,73.3, 60.6, 49.5, 49.0, 39.5, 33.6, 28.2, 27.9, 26.7, 25.6.Step D

The solution of the ester-acetal (4.31 g, 14.9 mmol) from the previousstep in dichloromethane (4 mL) was treated with trifluoroacetic acid(4.0 mL) and stirred at room temperature overnight. The solvent wasevaporated in vacuo, and the residue was co distilled several times withhexane to yield 4.14 g of the desired acid. ¹H NMR (500 MHz, CDCl₃):2.84 (d, J=18.31 Hz), 2.26 (d, J=18.76 Hz), 2.48 to 2.28 (m, 4H), 1.41(s, 3H), 1.37 (s, 3H).

Intermediate 12

A solution of Intermediate 11 (2.00 g, 10.7 mmol),3,5-bistrifluoromethylbenzyl

3.00 g, 10.7 mmol), 1-hydroxy-7-azabenzotriazole (1.46 mg, 10.7 mmol)and

pylamine (1.87 mL, 10.7 mmol) in dichloromethane (10 mL) was treatedwith 1-[3-dimethylamino)propyl]-3-ethylcarbodiimide EDC (3.09 g, 16.1mmol) and the reaction mixture was stirred at room temperatureovernight. It was diluted with dichloromethane, washed with water, driedwith anhydrous magnesium sulfate and the solvent was removed in vacuo.The crude residue was further purified by column chromatography (Silicagel, ethyl acetate:hexanes/4:1) to yield 1.52 g (36%) of the pureproduct. ¹H NMR (500 MHz, CDCl₃): 8.10 (bt, J=5.72 Hz, 1H), 7.77 (s,1H), 7.75 (s, 2H), 4.61 (dd, J=15.56, 6.18.Hz, 1H), 4.56 (dd, J=6.18 Hz,15.33 Hz, 1H), 2.86 (d, J=18.07 Hz, 1H), 2.40 to 2.18 (m, 4H), 1.32 (s,3H), 1.25 (s, 3H). ¹³C NMR (125 MHz, CDCl₃): 216.6, 175.8, 141.6, 131.9,131.7, 127.5, 121.9, 73.8, 58.5, 44.6, 42.7, 36.8, 28.5, 27.2, 26.5.LC-MS for C18H19F6NO3 [M+H]⁺ calc. 412.13, found 412.15.

Intermediate 13

Step A

The cis-racemate of 3-methyl-4-amino-tetrahydropyrane was obtained from3-methyltetrahydropyran-4-one (Intermediate 5) in a procedure analogousto that described under preparation of Intermediate 2.Step B

A solution of the racemic cis-amine (1.54 g, 10.3 mmol), the preparationof which is described in the previous step, and diisopropylethylamine(4.46 mL, 25.6 mmol) in dry dichloromethane, under N₂ at ambienttemperature, was treated with neat carbobenzoxy chloride (1.61 mL, 11.3mmol) and the resulting mixture was stirred at room temperature for 2 h.It was diluted with dichloromethane and extracted with 10% aqueoussolution of citric acid. The aqueous phase was back extracted withdichloromethane, and the combined organic extracts were washed withsaturated aqueous sodium bicarbonate. After drying (anhydrous magnesiumsulfate), the solvent was removed in vacuo and column chromatography(Silica gel, ethyl acetate:hexane/2:3) gave 1.8347 g (72%) of the pureproduct. The respective enantiomers were obtained by chiral HPLC using aChiralPak AD semi-preparative column. The absolute configuration of thefaster eluting isomer (Tr=13.0 minutes, Hexane:EtOH/93:7, 9 mL/min) wasshown to be (3R,4S) by both derivatization of the free amine followed byNMR spectroscopy, as well as single crystal X-ray diffraction analysis.¹H NMR (500 MHz, CDCl₃): 7.47 (bm, 5H), 5.12 (bs, 2H), 4.65 (bd, J=8.7Hz, 1H), 3.98 (dd, J=11.44, 3.43 Hz, 1H), 3.87 (dd, J=11.4, 4.3 Hz, 1H),3.45 (m, 2H), 3.08 (t, J=11.40 Hz, 1H), 1.95 (d, J=11.60 Hz, 1H), 1.50(m, 2H), 0.90 (d, J=6.63 Hz, 3H).Step C

The solution of the CBZ-protected amine from the previous step (284 mg,1.14 mmol) in ethanol (15 mL) was hydrogenated using 133 mg of Pd/C(10%) under an ambient hydrogen pressure of a balloon for 30 minutes.The catalyst was filtered off, and the solution was concentrated invacuo to leave 158 mg (91%) of the desired product.

Intermediate 14

Step A

A solution of the acid Intermediate 9 (3.72 g, 13.0 mmol),3,5-bistrifluoromethylbenzylamine hydrochloride (3.62 g, 13.0 mmol),diisopropylethylamine (2.26 mL, 13.0 mmol), 1-hydrozy-7-azabenzotriazole(1.76 g, 13.0 mmol) in dichloromethane (30 mL) was treated with EDC(3.72 g, 19.4 mmol) and the reaction mixture was stirred at roomtemperature for 2 h. It was poured onto water (50 mL) and extracted withdichloromethane. The combined organic extracts were washed with brine,dried with anhydrous. magnesium sulfate and the solvent was removed invacuo to leave 4.80 g of an oily crude product. This was furtherpurified by column chromatography (Silica gel, ethyl acetatehexanes/2:3) to yield 3.18 g (48%) of the pure product. ¹H NMR (500 MHz,CDCl₃): 8.40 (bs, 1H), 7.76 (s, 1H), 7.75 (s, 2H), 5.34 (d, J=6.18 Hz,1H), 4.56 (m, 2H), 4.0 (m, 1H), 3.21 (s, 1H), 2.15 (dd, J=14.2, 4.81 Hz,1H), 2.05 to 1.85 (m, 4H), 1.62 (m, 1H), 1.41 (bs, 9H), 1.26 (s, 3H),1.23 (s, 3H). ¹³C NMR (125 MHz, CDCl₃): 178.4, 155.7, 141.8, 131.9 (m),127.5, 121.0, 79.1, 74.6, 52.3, 42.7, 37.8, 33.4, 31.6, 28.3, 27.0,26.3.Step B

The solution of the BOC-protected amine from the previous step (3.18 g,6.20 mmol) was stirred at room temperature in dioxane/HCl (4.0 N) for 1h. The solvent was removed in vacuo to yield the pure hydrochloride(2.63 g, 94%). LC MS for C₁₈H₂₂F₆N₂O₂ for [M+H]⁺ calc. 413.16, found413.20.

Intermediate 15

Step A

A solution of tetrahydro-4H-pyran-4-one (10.0 g, 99.9 mmol) in drytetrahydrofuran (200 mL) was cooled to 0° C. and a solution ofmethylmagnesium chloride (3.0 M, in THF) was added via syringe. Afterstirring at 0° C. for 30 minutes the cooling bath was removed, and thereaction was allowed to proceed at room temperature for another 30minutes. It was quenched by pouring onto a saturated aqueous solution ofammonium chloride, and extracted with diethyl ether. After drying withmagnesium sulfate the solvent was removed in vacuo (100 mmHg), and thecrude product (11.07 g) was further purified by distillation (b.p.: 87°C. @ 20 mmHg) to yield 4.36 g (38%) of the desired alcohol. ¹H NMR (500MHz, CDCl₃): 3.75 (ddd, J=14.2, 11.2, 2.8 Hz, 2H), 3.65 (dt, J=8.7, 4.4Hz, 2H), 1.98 (bs, 1H), 1.63 (ddd, J=14.2, 10.3, 4.6 Hz, 2H), 1.50 (bd,J=13.0 Hz, 2H), 1.24 (s, 3H).Step B

A solution of 4-hydroxy-4-methyltetrahydropyran (5.86 g, 50.5 mmol) inacetonitrile (25 mL) was cooled to 0° C. and concentrated sulfuric acid(10.5 g) was added. The reaction mixture was stirred at room temperaturefor another 1 h, after which time it was poured onto ice (˜50 g) andextracted with dichloromethane. The combined organic phases were washedwith a saturated solution of sodium bicarbonate, dried with anhydrousmagnesium sulfate, and the solvent was evaporated to dryness in vacuo.The crude product (7.15 g) was dissolved in a solution consisting of 20g of sodium hydroxide and 30 mL of ethylene glycol in 20 mL of water,and was heated to 125° C. for 72 h. The reaction mixture was allowed tocool to room temperature, and the pH was made strongly acidic withconcentrated sulfuric acid (˜70 mL). The entire mixture was evaporatedto dryness under reduced pressure and allowed to dry at high vacuumovernight. The amine was liberated from the sulfate form with aqueoussodium hydroxide, and extracted into dichloromethane. The combinedextracts were dried with anhydrous sodium sulfate, and thedichloromethane was evaporated to dryness in vacuo. The pure amine wasobtained by distillation at reduced pressure: b.p.: 90-91° C. @ 100mmHg; 3.3983 g (59%) of the product was obtained in a form of a mobileliquid. ¹H NMR (500 MHz, CDCl₃): 3.71 (ddd, J=11.44, 8.01, 3.20 Hz, 2H),3.58 (ddd, J=10.52, 6.40, 3.89 Hz, 2H), 1.56 (bs, 2H), 1.36 (m, 2H),1.12 (s, 3H). ¹³C NMR (125 MHz, CDCl₃): δ4.3, 46.3, 40.6, 27.0.

Intermediate 16

Step A

To a stirred solution of Intermediate 10 (2.09 g, 7.71 mmol),1-ethyl-3-(3-dimethyl-aminopropyl)carbodiimide (2.96 g, 15.4 mmol) inDCM (100 mL), was added 3,5-bistrifluorobenzylamine hydrochloride (2.26g, 8.10 mmol), diisopropylethylamine (1.05 g, 8.10 mmol), and1-hydroxy-7-azabenzotriazole (1.15 g, 8.48 mmol). The reaction wasstirred at room temperature for 18 h before being diluted with DCM andwashed twice with aqueous 1 N HCl, once with saturated aqueous sodiumbicarbonate, and once with brine. The organic layer was dried overMgSO₄, filtered and concentrated under reduced pressure. The product waspurified by medium pressure liquid chromatography (silica gel, 60%EA/Hexanes) to give 2.23 g of a colorless oil which was used directly inStep B.Step B

The product from Step A was dissolved in hydrogen chloride (4 N solutionin dioxane, 25 mL) and stirred at room temperature. After 1.5 h thereaction was concentrated under reduced pressure to give 1.79 g of awhite solid (54% over 2 steps). ESI-MS calc. for C18H22F6N2O: 396.4;found 397.2 (M+H).

Intermediate 17

Step A

To a cooled (0° C.) solution of Intermediate 10 (1.0 g, 3.7 mmol) inmethanol (50 mL) was added thionyl chloride (1.1 mL, 15 mmol) dropwiseand the resulting solution was allowed to warm to room temperature.After 18 h, an additional amount of thionyl chloride (2.1 mL, 30 mmol)was added and the reaction was allowed to stir at room temperature.After 18 h, the reaction was concentrated under reduced pressure and theproduct was used directly in Step B.Step B

The crude product from Step A was combined withtetrahydro-4H-pyran-4-one (730 mg, 7.3 mmol), triethylamine (1.0 mL, 7.3mmol), 4 Å powdered molecular sieves (˜1 g), and sodiumtriacetoxyborohydride (5.1 g, 24 mmol) in 50 mL DCM. The reactionmixture was stirred at room temperature for 4 days, then filteredthrough celite, diluted with DCM, and washed with saturated NaHCO₃solution twice and then once with brine. The organic layer was driedover anhydrous MgSO₄, filtered and concentrated to give 1.0 g of aslightly yellow oil (99+% over 2 steps). ESI-MS calc. for C15H27NO3:269, found 270 (M+H).Step C

The product from Step B (1.0 g, 3.7 mmol) was combined with formaldehyde(37% solution in water) (3.0 mL, 37 mmol), 4 Å powdered molecular sieves(1 g), and sodium triacetoxyborohydride (3.9 g, 19 mmol) in 50 mL DCM.The reaction mixture was stirred at room temperature for 1 h, thenfiltered through celite, diluted with DCM, and washed with saturatedNaHCO₃ and brine. The organic layer was dried over anhydrous MgSO₄,filtered and concentrated to give 1.0 g of a slightly yellow oil (95%).ESI-MS calc. for C16H29NO3: 283; found 284 (M+H).Step D

The product from Step C (1.0 g, 3.5 mmol) was dissolved in a solution ofTHF (10 mL) and methanol (10 mL) and treated dropwise with a solution oflithium hydroxide monohydrate (730 mg, 17 mmol) in water (10 mL) over 10min. The reaction was allowed to stir at room temperature for 1 h beforebeing heated to reflux. After 36 h at reflux the reaction was cooled toroom temperature, neutralized with 3N hydrochloric acid and concentratedunder reduced pressure to dryness. The resultant crude product wastriturated with a 50% solution of methanol/DCM to give 950 mg of a whitesolid (99%). ESI-MS calc. for C15H27NO3: 269: found 270 (M+H).

Intermediate 18

Step A

A mixture of tetrahydro-4H-pyran-4-one (10 g, 100 mmol) and pyrrolidine(11 g, 150 mmol) was stirred at room temperature for 1 h. The excesspyrrolidine was removed on the vacuum pump and the residue was driedunder high vacuum overnight. The enamine was obtained as a yellow liquid(14.7 g) which was used in next step without further purification.Step B

The enamine from step A (1.54 g, 10.0 mmol) and 4-N,N-dimethylpyridine(1.22 g) were treated with DMF (25 mL). The mixture was cooled to 0° C.and solid 5-(trifluoromethyl)dibenzo-thiopheniumtrifluoromethanesulfonate (4.02 g, 10.0 mmol) was added. The resultingmixture was stirred at 0° C. for 1 h, quenched with 30 mL ofconcentrated aqueous HCl, stirred for 2 h, and extracted with ether(4×70 mL). The combined ether layers were washed with water (50 mL) andbrine (50 mL), dried over Na₂SO₄, evaporated. The residue was purifiedby flash chromatography on silica gel (10% ethyl ether/Hexane) to yieldtwo components. The more polar component (200 mg) was the desiredmaterial. 1H-NMR showed it might exist in semi ketal form. ¹H NMR (400,CDCl₃): 4.43-3.38 (m, 5H), 3.24, 3.18 (ss, 3H), 2.52 (m, 1H), 1.82 (m,1H). The less polar product (100 mg) was confirmed as alpha-alpha′di-trifluoromethyl tetrahydro-2H-pyran-4-one. ¹H NMR (400 MHz, CDCl₃):4.59 (dd, 2H), 3.24, 3.80 (t, J=11.3, 2H), 3.42 (m, 2H).

Intermediate 19

To a cooled (−78° C.) solution of lithium diisopropylamide (2.0 Msolution in heptane/THF/ethylbenzene, 65 mL, 200 mmol) in THF (300 mL)was added trimethylsilyl chloride (70 mL, 500 mmol) dropwise. After 5minutes at −78° C. tetrahydro-4H-pyran-4-one (10 g, 100 mmol) in THF(120 mL) was added. The reaction was stirred for 1 minutes beforetriethylamine (200 mL) was added and the reaction was quenched withsaturated aqueous sodium bicarbonate. The solution was extracted twicewith diethyl ether and the combined organic layers where washed with 0.1N aqueous citric acid, dried over K₂CO₃, filtered and concentrated underreduced pressure. The resultant silyl ether was dissolved in THF (90 mL)and cooled to 0° C. N-bromosuccinimide (19.6 g, 110 mols) was addedportionwise and the resulting solution was stirred 5 minutes at 0° C.The ice bath was removed and the solution was stirred for 30 minutes atroom temperature before being quenched with saturated aqueous sodiumbicarbonate and extracted twice with diethyl ether. The combined organiclayers where washed with brine, dried over MgSO₄, filtered andconcentrated under reduced pressure. The product was purified by flashchromatography (silica gel, 20-50% diethyl ether/petroleum ether) togive 8.75 g of a white solid (49%).

H NMR (CDCl₃, 500 MHz): 4.46 (t, J=6.5 Hz, 1H), 4.30 (m, 1H), 4.09 (m,1H), 3.91 (m, 2H), 2.99 (m, 1H), 2.65 (m, 1H).

Intermediate 20

Step A

This intermediate was prepared in an analogous fashion to that ofIntermediate 10, except 2,4,6-triisopropylbenzene-sulfonylazide was usedin place of 2-iodopropane. The cis and trans isomers were not separated;therefore, the compound was used as a mixture of two diastereomers.FT-IR: 3310, 2939, 2568, 2110, 1711, 1680.Step B

A mixture of the acid described in step A, Intermediate 20 (250 mg, 0.93mmol), 3,5-bis(trifluoromethyl)benzylamine hydrochloride (260 mg, 0.93mmol), DMAP (12.7 mg, 0.093 mmol), N,N-diisopropylethylamine (160 □l,0.93 mmol) in dichloromethane (20 mL) was treated with1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EDC, 356mg, 1.86 mmol) and stirred at room temperature overnight. The reactionmixture was diluted with dichloromethane (20 mL), washed with water(2×20 mL), brine (1×30 mL), dried over anhydrous sodium sulfate and thesolvent was evaporated. Purification by preparative TLC (eluant: 25%ethyl acetate/75% hexane) afforded two separate single isomers (isomer1, less polar cis, 72 mg, 18%; isomer 2, more polar trans, 140 mg, 31%).LC-MS calculated for C₂₀H₂₃F₆N₅O₃ is 495.17, found (MH)⁺ 495.2 for bothisomer 1 and isomer 2. FT-IR still exhibited 2109 cm⁻ for the azide. ¹HNMR (CDCl₃, 500 MHz) (for isomer 1): 7.84 (s, 1H), 7.72 (s, 2H), 7.15(br s, 1H), 5.58 (br s, 1H), 4.61 (dd, J=6.0, 15.56 Hz, 1H), 4.58 (dd,J=6.0, 15.56 Hz, 1H), 4.37 (br s, 1H), 2.44 (ddd, J=8.1, 8.9, 13.8 Hz,1H), 2.34 (dd, J=7.3, 14.6 Hz, 1H), 2.24 (dd, J=4.6, 14.5 Hz, 1H),2.16-2.02 (m, 2H), 1.96-1.88 (m, 1H), 1.46 (s, 9H).

¹H NMR (CDCl₃, 500 MHz) (for isomer 2): 7.83 (s, 1H), 7.73 (s, 2H), 7.02(br s, 1H), 4.67 (br s, 1H), 4.62 (dd, J=6.2, 15.3 Hz, 1H), 4.57 (dd,J=6.0, 15.3 Hz, 1H), 4.27 (br s, 1H), 2.68 (dd, J=8.0, 14.5 Hz, 1H),2.40-2.29 (m, 2H), 2.11-2.03 (m, 1H), 1.92 (br d, J=12.1 Hz, 1H),1.73-1.65 (m, 1H), 1.44 (s, 9H).Step C

To a solution of the less polar cis isomer (isomer 1) described in stepB, intermediate 20 (70 mg, 0.141 mmol) in THF/water (2 mL/0.1 mL) wasadded triphenyl-phosphine (111 mg, 0.423 mmol) and the resultingsolution stirred for 5 h at room temperature. The reaction wasevaporated under reduced pressure and the residue was purified bypreparative TLC (eluant: 90% ethyl acetate/10% hexane) to affordIntermediate 20 (50 mg, 60%) as a yellow foam. LC-MS calculated forC₂₀H₂₅F₆N₃O₃ is 469.18, found (MH)⁺ 470.2

Intermediate 21

A mixture of 9.70 g (0.0970 mol) of tetrhydro-4H-pyran-4-one and 10.5 g(150 mmol) of pyrrolidine was stirred at room temperature for 1.5 h. Theexcess of pyrrolidine was removed on vacuum pump. The residue wasdissolved in 90 mL of ether, cooled to 0° C. and 7.4 mL of acrolein wasadded. The resulting mixture was stirred at room temperature overnight.67 mL of water was added, followed by a solution of 14 g of sulfuricacid (98%) in 33 mL of water. The ether and 10 mL of water were removedunder reduced pressure, the remaining mixture was refluxed for 0.3 h andthen cooled to room temperature. The resulting dark mixture wasextracted with DCM (4×100 mL), and the combined organics where driedover anhydrous Na₂SO₄ and evaporated. The residue was purified on MPLC(30% ethyl acetate/hexane). A mixture (6.6 g) of endo/exo isomers (˜1/1)was obtained together with pure fast isomer (1.0 g, endo) and pure slowisomer (0.8 g, exo). ¹H NMR (400 MHz, CDCl₃): endo: 4.58 (d, J=11.6 Hz,1H), 4.20 (d, J=11.2 Hz, 1H), 4.17 (d, J=11.2 Hz, 1H), 3.91 (d, J=11.3Hz, 1H), 3.72 (d, J=11.5 Hz, 1H), 2.60-2.30 (m, 4H), 2.13 (m, 1H), 2.02(m, 1H), 1.80 (m, 1H). Exo: 4.54 (d, J=1.1 Hz, 1H), 4.10 (dd, J=11.4 Hz,2H), 3.80 (dd, J=11.5 Hz, 2H), 2.86 (s, 1H), 2.70 (m, 1H), 2.50 (s, 1H),2.38 (m, 2H), 2.10 (m, 1H), 1.78 (m, 1H).

Intermediate 22

Step A

To a mixture of Intermediate 21 (endo/exo: ˜1:1, 3.12 g, 20 mmol) andDBU (9.0 g, 60 mmol) in benzene (25 mL) at 0° C. was added dropwise aneat solution of trifluoromethane sulfonic acid anhydride. An exothermicreaction was observed. The reaction mixture was stirred for 1 h, dumpedonto a silica gel column, eluted with 20% Et₂O/hexane. The desiredproduct was obtained as a light yellow oil (1.80 g).). ¹H NMR (400 MHz,CDCl₃): 5.98 (m, 1H), 5.65 (m, 1H), 4.10 (dd, 1H), 3.90(dd, 1H), 3.78(dd, 1H), 3.65 (dd, 1H), 2.80 (m, 3H), 2.50 (d, J=11.5).Step B

A mixture of the Intermediate 21 (9.0 g) and 10% Pd/C (0.9 g) in 50 mLof ethyl acetate was hydrogenated on a Parr-shaker for 2 h under 50 psiof hydrogen. The catalyst was removed by filtration. The filtrate wasevaporated. Intermediate 22 was obtained as a light yellow solid (6.817g). ¹H NMR (400 MHz, CDCl₃): 4.24 (d, J=11.5 Hz, 2H), 3.90 (d, J=11.60Hz, 2H), 2.58 (m, 1H), 2.38 (br S, 2H), 2.25 (m, 2H), 2.08 (m, 2H), 1.58(m, 1H).

Intermediate 23

Step A

To a flame-dried, 125 mL flask under nitrogen was placeddiisopropylamine (4.7 g, 33 mmol) and 20 mL of anhydrous THF at −78° C.A solution of n-butyllithium (1.6 M, 20.5 mL, 33 mmol) was added. Thecooling bath was removed and the reaction mixture was warmed to roomtemperature for 10 minutes, then recooled to −78° C. A solution oftrimethylsilyl chloride (10 mL, 150 mmol) in 20 mL of THF at −78° C. wascannulated into the reaction flask. Then a solution of the Intermediate5 (3.5 g, 30 mmol) in 20 mL of THF was cannulated into the reaction. Alarge amount of white precipitate was formed. 30 mL of triethylamine wasadded. The reaction mixture was quenched with saturated aqueous sodiumbicarbonate and extracted with pentane (2×250 mL). The pentane layerswere washed with water (250 mL), 5% aqueous citric acid (2×250 mL) anddried over anhydrous sodium sulfate. The crude product was used in nextstep without further purification. ¹H NMR (400 MHz, CDCl₃): 4.74 (s,1H), 4.18 (s, 2H), 3.84 (dd, J=4.6 Hz, 1H), 3.42 (dd, J=4.5 Hz, 1H),2.20 (m, 1H), 1.03 (d, J=7.0 Hz, 3H), 0.19 (s, 9H).Step B

Diflurotriethylenediammonium tetrafluroborate (10.6 g, 30.0 mmol) wasplaced in a 100 mL flask under nitrogen. 50 mL of anhydrous acetonitrilewas added. The mixture was stirred at 0° C., then a neat solution of theenol trimethylsilyl ether (entire material from step A, intermediate 23)was added dropwise. The reaction was stirred until all the enol etherwas consumed (˜30 min). Acetonitrile was then removed and the residualsolid was washed with a mixture of ether and hexane (1/9, 5×). Thefiltrates were evaporated and the residue was purified on MPLC. Twocomponents were obtained. The proton NMR of the slow-eluted material wasconsistent with that of the desired product (250 mg). ¹H NMR (400 MHz,CDCl₃): 4.79; 4.67 (m, 1H), 4.19 (m, 1H), 4.08 (m, 1H), 3.84 (m, H),3.40 (t, J=1.0 Hz, 1H), 3.16 (m, 1H), 1.06 (d, J=6.9 Hz, 3H). Thefast-eluted compound (200 mg) was alpha-alpha′ difluoro ketone.

Intermediate 24

Step A

To a cooled (0° C.) mixture of 5,6-dihydro-4-methoxy-2H-pyran (10.0 g,87.5 mmol) in methanol (200 mL) was added dropwise a solution of m-CPBA(30.2 g, 175 mmol) in methanol (50 mL) via addition funnel. Theresulting solution was stirred for 5 h allowing to warm to roomtemperature. The methanol was removed under reduced pressure affording awhite solid. The material was dissolved in 500 mL of dichloromethane andcooled to 0° C. To the mixture, while stirring vigorously, was added inportions an excess of solid calcium hydroxide (50-60 grams). Afterstirring an additional 30 minutes, the mixture was filtered through aplug of celite and the filtrate evaporated under reduced pressure toafford 11.62 g (82%) of the desired product as a clear oil. ¹H NMR(CDCl₃, 500 MHz) δ 3.88-3.80 (m, 2H), 3.73-3.68 (m, 2H), 3.54-3.48 (m,1H), 3.28 (s, 3H), 3.27 (s, 3H), 2.00-1.93 (m, 1H), 1.82-1.77 (m, 1H).Step B

To a cooled (0° C.) solution of the product from Step A (9.40 g, 58.0mmol) in THF (200 mL), under nitrogen, was slowly added NaH (2.32 g,58.0 mmol) and the resulting slurry was stirred for 1 h at 0° C.Iodomethane (7.22 mL, 116 mmol) was then added via syringe to the slurryand the resulting mixture was stirred overnight allowing to warm to roomtemperature. The reaction was quenched with a saturated solution ofammonium chloride (200 mL) and the organic layer was then removed usinga separatory funnel. The aqueous layer was extracted with ether (3×150mL) and all the organics were combined, dried over anhydrous sodiumsulfate, filtered, and evaporated in vacuo. Purification was done byflash column using a stepwise gradient eluant of 10-60% ether/hexane toafford 8.46 g (83%) of the desired product as a clear oil. ¹H NMR(CDCl₃, 500 MHz) 3.98 (dd, J=2.5, 12.4 Hz, 1H), 3.77 (ddd, J=3.5, 7.1,10.8 Hz, 1H), 3.57 (dd, J=1.4, 12.4 Hz, 1H), 3.50 (dd, J=2.5, 11.7 Hz,1H), 3.46 (s, 3H), 3.25 (s, 3H), 3.22 (s, 3H), 3.22-3.20 (m, 1H), 1.96(ddd, J=4.7, 11.8, 16.5 Hz, 1H), 1.75 (br dd, J=1.7, 14.2 Hz, 1H).Step C

A solution of product from step B, intermediate 24 (3.0 g, 17 mmol) inTHF/water (60 mL/10 mL) was treated with concentrated hydrochloric acid(6 mL) and the resulting solution stirred at room temperature for 1 h.The mixture was concentrated in vacuo to remove the THF and the aqueouslayer then extracted with ether (6×50 mL). The organics were combined,dried over anhydrous sodium sulfate, filtered, and evaporated underreduced pressure to afford intermediate 24 (1.75 g, 79%) as a clear oil.¹H NMR (CDCl₃, 500 MHz) 4.23 (ddd, J=1.2, 11.4, 12.4 Hz, 1H), 4.15-4.09(m, 1H), 3.82 (dd, J=5.95, 8.7 Hz, 1H), 3.74 (ddd, J=5.5, 8.5, 13.6 Hz,1H), 3.56 (dd, J=8.8, 11.3 Hz, 1H), 3.50 (s, 3H), 2.61 (app dd, J=5.0,8.9 Hz, 2H).

Intermediate 25

Procedure A

To a solution of terahydro-4H-pyran-4-one (700 mg, 7.00 mmol) and HMPA(1.2 mL) in THF (14 mL) was added slowly a solution of LDA (3.5 mL, 2 Msolution) in 14 mL THF at −78° C. The mixture was stirred for 5 minutesbefore iodoethane (0.56 mL, 7.0 mmol) was added and the solution wasgradually warned to 0° C. over 2 h. The reaction mixture was quenchedwith a saturated solution of NH₄Cl and then extracted with ether (4×50mL). The ether layer was washed with brine, dried over anhydrousmagnesium sulfate, concentrated and purified by flash columnchromatography. Eluting with hexane:EtOAc (19:1) afforded intermediate25 (0.07 g, 8%).

Procedure B

Step A

A mixture of ethyl 2-ethylacetoacetate (3.0 g, 19 mmol), ethylene glycol(1.4 g, 23 nmol), CSA (100 mg) and benzene (80 mL) was refluxed in aDean-Stark apparatus, with continuos removal of water. After ensuringthe completion of the reaction (by TLC) it was diluted with water andextracted with ether (100 mL). The ether layer was washed with brine,dried (anhydrous magnesium sulfate) and concentrated to afford thedesired compound (4.2 g). This was taken in ether (50 mL) and was slowlyadded to LAH (1.2 g, 32 mmol) at 0° C. The reaction was warmed to roomtemperature and stirred for 12 h. The reaction mixture was then quenchedsequentially with water (1.5 mL), 15% NaOH (1.5 mL) and water (4.5 mL).The resultant heterogeneous mixture was vigorously stirred and filtered.Evaporation of the filtrate gave 3.1 g of the title compound thatrequired no further purification.Step B

To a stirring slurry of silica (12 g, 230-400 mesh) in methylenechloride (100 mL) was added a 10% aqueous solution of oxalic acidfollowed by the acetal from Step A (1.6 g, 10 mmol) in methylenechloride (5 mL). The resultant mixture was stirred at room temperatureuntil the reaction was complete. Upon the completion of the reaction,NaHCO₃ (1.0 g) was added, stirred (10 minutes) and filtered. Thefiltrate was evaporated to give the 0.96 g of the title compound thatrequired no purification.Step C

To a premixed solution of triethyl orthoformate (2.4 g, 16 mmol) and tin(IV) chloride (16.3 mL 1.0 M solution in dichloromethane, 16 mmol) at40° C. was added the hydroxy ketone from Step B (0.95 g, 8.1 mmol) indichloromethane (3 mL). The reaction mixture was warmed to −5° C. over1.5 h before being quenched with a saturated NaHCO₃ solution andextracted with ether (2×50 mL). The ether layer was washed with brine,dried over anhydrous magnesium sulfate, concentrated and purified byflash column chromatography on silica gel. Eluting with hexane:ether(9:1) to give the title compound (0.58 g, 57%).Step D

The intermediate from Step C (0.56 g) in hexane (10 mL) and Pd/C (5%, 50mg) was hydrogenated at room temperature until the TLC indicated thecompletion of reaction. The reaction mixture was filtered and thefiltrate was carefully evaporated (volatile product!) to yield themixtures of the desired intermediate 25 and the over reduction product.The recovery of intermediate 25 was further facilitated by a subsequentTPAP/NMMO/DCM oxidation of the mixture, which after 1 h was filtered toyield 410 mg of the title compound that required no furtherpurification.

1H NMR (CDCl₃, 500 MHz): 4.15 (m, 2H), 3.80 (m, 1H), 3.48 (m, 1H), 2.61(m, 1H), 2.44 (m, 2H), 1.82 (m, 1H), 1.31 (s, 1H), 0.72 (m, 1H), 0.94(t, J=7.4 Hz, 3H).

Intermediate 26

To a solution of terahydro-4H-pyran-4-one (700 mg, 7.0 mmol) and HMPA(1.2 mL) in THF (14 mL) was added slowly a solution of LDA (3.5 mL, 2 Msolution) in 14 mL THF at −78° C. The mixture was stirred for 5 minutesprior to adding n-iodopropane (0.58 mL, 7.0 mmol). The reaction mixturewas gradually warmed to 0° C. over 3 h before it was quenched with asaturated solution of NH₄Cl and then extracted with ether (4×50 mL). Theether layer was washed with brine, dried (anhydrous magnesium sulfate),concentrated and purified by flash column chromatography. Eluting withhexane:EtOAc (19:1) gave intermediate 26 (0.07 g, 7%).

Intermediate 27

Following the Steps A-D given for the preparation of Intermediate 25(Procedure 2) and starting from methyl 2,2-dimethylacetoacetate, gavethe title compound. ¹H NMR (CDCl₃, 500 MHz): 3.98 (m, 2H), 3.58 (s, 2H),2.56 (m, 2H), 1.15 (s, 6H).

Intermediate 28

Following the Steps A-D given for the preparation of Intermediate 25(procedure 2) and starting from methyl 2,4-dimethyl-3-oxobutyrate, gavethe title compound. ¹H NMR (CDCl₃, 500 MHz): 4.22 (m, 1H), 3.99 (m, 1H),3.62 (m, 1H), 3.28 (m, 1H), 2.72 (m, 1H), 1.16 (d, J=6.8 Hz, 3H), 0.97(d, J=6.8 Hz, 3H).

Intermediate 29

To a mixture of 5.6-dihydro-4-methoxy-2H-pyran (500 mg, 4.4 mmol) inacetonitrile/water (15 mL, 1:1) at room temperature was added[1-(chloromethyl)-4-fluoro-1,4-diazoniabicyclo[2.2.2.] octanebis(tetrafluoroborate)] (1.5 g, 4.4 mmol, SELECTFLUOR™) in one lot andthe reaction stirred to completion. Solid NaCl was added to the reactionmixture, and then extracted with ether (4×50 mL). The ether layer wasdried (anhydrous magnesium sulfate) and concentrated to yield 0.34 g(65%) of the title compound that required no further purification. ¹HNMR (CDCl₃, 500 MHz): 4.95 (m, 1H), 4.4-4.21 (m, 2H), 3.72-3.65 (m, 2H),2.75 (m, 2H).

Intermediate 30

Step A

To a mixture of 5.6-dihydro-4-methoxy-2H-pyran (2.0 g, 18 mmol) inmethanol (40 mL) at 0° C. was added m-CPBA (6.0 g, 35 mmol). Afterstirring for 10 min. at 0° C., the reaction mixture was allowed to warmto room temperature and stirred for 1 h. The solvent was removed undervacuum and the crude mixture was chromatographed on a silica column.Eluting with hexane:EtOAc (7:3) gave 2.8 g (95%) of the title compound.

1H NMR (CDCl₃, 500 MHz): 3.83 (m, 2H), 3.70 (m, 2H), 3.50 (m, 1H), 3.28(s, 3H), 3.27 (s, 3H), 1.96 (m, 1H), 1.77 (m, 1H).Step B

To a mixture of the acetal from Step A (2.8 g, 17 mmol) indichloromethane (30 mL) was added 4 Å powdered molecular sieves (˜5 g),4-methylmorpholine N-oxide (5.0 g, 43 mmol) and finally TPAP (0.2 g).The resultant mixture was stirred vigorously for 3 h at which point thereaction was complete. It was filtered, evaporated, and purified byflash column chromatography on silica gel. Eluting with hexane:ether(1:4) gave the title compound (2.67 g, 96%).Step C

To the ketone from Step B (2.8 g, 18 mmol) in THF (20 mL) at 0° C. wasadded TBAF (28 mg) followed by neat trimethyl(trifluoromethyl)silane(4.0 g, 28 mmol). After stirring for 10 min. at 0° C., the reactionmixture was allowed to warm to room temperature and stirred for 12 h.THF was removed under vacuum and the crude passed through a silica gelcolumn. Eluting with hexane:EtOAc (4:1) gave 4.1 g (77%) of the titlecompound.Step D

To the silylether from Step C (4.0 g, 13.3 mmol) at room temperature wasadded TFA (2.0 mL) and the mixture was stirred for 36 h. The TFA wasremoved under vacuum and the crude was purified by flash silica gelcolumn chromatography. Eluting with hexane:ether (4:1) gave 1.5 g (65%)of the title compound.

1H NMR (CDCl₃, 500 MHz): 4.49 (m, 1H), 4.41 (m, 1H), 4.34 (br, 1H), 3.72(m, 1H), 3.39 (m, 1H), 3.07 (m, 1H), 2.72 (m, 1H).

Intermediate 31

Prepared according to J. Am. Chem. Soc., 1997, 119, 4285, except thatthe reaction was performed on the ethyl ester.

Intermediate 32

Step A

To a solution of trans-1-methoxy-3 (trimethylsilyloxy)-1,3-butadiene(1.62 g, 9.4 mmol) and acetaldehyde (0.63 mL, 11 mmol) in ether (25 mL)at −78° C. was added slowly a solution of boron trifluoridediethyletherate (1.36 mL, 10.7 mmol). After stirring for 3 h at the sametemperature, the reaction mixture was quenched with a saturated solutionof sodium bicarbonate, warmed to room temperature and then extractedwith ether (2×50 mL). The ether layer was washed with brine, dried(anhydrous magnesium sulfate), and concentrated followed by purificationby silica flash column chromatography. Eluting with hexane:diethyl ether(8:2) gave the desired product (0.6 g, 57%).Step B

The intermediate from Step A (0.59 g) in EtOAc (10 mL) with Pd/C (50 mg)was hydrogenated at room temperature under a hydrogen filled balloon,until the reaction was complete as indicated by TLC. The reactionmixture was filtered and the filtrate was carefully concentrated(volatile product!) followed by purification by silica gel flash columnchromatography. Elution with hexane:diethyl ether (7:3) gave 0.2 g (33%)of the desired compound (Intermediate 30).

1H NMR (CDCl₃, 500 MHz): 4.28 (m, 1H), 3.75 (m, 1H), 3.69 (m, 1H),2.63-2.27 (m, 4H), 1.34 (d, J=6.1 Hz, 3H).

Intermediate 33

Step A

To comanic acid (0.25 g) in EtOAc (5 mL) was added Pd/C (25 mg) and theresulting solution was hydrogenated at room temperature under a hydrogenfilled balloon, until the reaction was complete as indicated by TLC. Thereaction mixture was filtered and the filtrate was concentrated to givethe desired product (0.25 g, 97%), which required no furtherpurification.Step B

To a solution of the intermediate from Step A (0.1 g, 0.7 mmol) inTHF:hexane (4 mL, 1:1) at room temperature, was added (trimethylsilyl)diazomethane (451 l, 1.3 eq, 2 M solution in hexane) and the mixture wasstirred for 24 h. The solvent was carefully concentrated (volatileproduct!) followed by purification by silica gel flash columnchromatography. Elution with hexane:EtOAc (4:1) gave the desired product(0.02 g, 16%).

Intermediate 34

Prepared according to Chem. Ber., 1959, 91, 1589.

Intermediate 35

Prepared according to Synlett, 1991, 783.

Intermediate 36

Prepared according to Synlett, 1991, 783.

Intermediate 37

Step A

To a solution of furan (20 mL, 720 mmol) in benzene (600 mL) was addedtetrabromoacetone (11 g, 30 mmol). The solution was cooled to 0° C. and1.0 M diethyl zinc solution in hexane (30 mL, 30 mmol) was addeddropwise. The solution was stirred at 0° C. for 2.5 h then at roomtemperature for 60 h. The reaction was quenched with a saturated aqueoussolution of dibasic EDTA (15 mL) and extracted with ethyl acetate (150mL). The organic layer was washed three times with saturated aqueousdibasic EDTA then with brine and was dried over MgSO₄, filtered andconcentrated under reduced pressure.Step B

The resultant oil from Step A was dissolved in a saturated solution ofNH₄Cl in methanol (120 mL) which was then added dropwise to Zn—Cu couple(20 g wet with diethyl ether). The mixture was stirred at roomtemperature for 2.5 h. The reaction was filtered through celite andconcentrated to remove the methanol. The product was dissolved in DCMand washed twice with saturated aqueous dibasic EDTA. The organic layerwas dried over MgSO₄, filtered and concentrated under reduced pressure.The product was purified by flash chromatography (silica, 33% PE/Et₂O)to give 1.7 g of a colorless solid (46%). ¹H NMR (CDCl₃, 500 MHz): 6.25(s, 2H), 5.03 (d, J=5.0 Hz, 2H), 2.75 (dd, J=5.0 Hz, 17.0 Hz, 2H), 2.32(d, J=16.5 Hz, 2H).Step C

To a solution of the product from Step B (780 mg, 6.4 mmol) in ethylacetate (25 mL) was added 78 mg of Pd(OH)₂ (20% on activated carbon)catalyst. A hydrogen balloon was placed over the reaction and it wasstirred at room temperature for 3 h. The reaction was filtered throughcelite, washed with ethyl acetate and concentrated to give 754 mg of acolorless oil (97%). ¹H NMR(CDCl₃, 500 MHz): 4.68 (bs, 2H), 2.65 (dd,J=5.0 Hz, 15.0 Hz, 2H), 2.23 (d, J=15.0 Hz, 2H), 2.20-1.98 (m, 2H),1.76-1.69 (m, 2H).

Intermediate 38

A solution of tetrahydro-4H-pyran-4-one (9.87 g, 98.5 mmol) and HMPA (18mL, 99 mmol) in THF (50 mL) was added dropwise under argon to a solutionof 1.5 M LDA.THF in cyclohexane diluted with THF (250 mL) at −78° C. Themixture was stirred for 20 min, then allyl bromide (17.0 mL, 197 mmol)in THF (50 mL) was added dropwise. The reaction mixture was warmed to 0°C. and stirred for 1.5 h. Then the reaction mixture was further warmedto room temperature and stirred for 30 min. The reaction was quenched bypouring into ice water. This mixture was extracted with ether threetimes. The combined ethereal layers were washed with brine, dried overMgSO₄, filtered, and concentrated. Purification by flash chromatography(silica, 35% ether/pet ether) provided 4.26 g of a colorless liquid(31%). ¹H NMR (CDCl₃, 400 MHz): 5.76 (m, 1H), 5.04-5.10 (m, 2H),4.16-4.22 (m, 2H), 3.77 (dt, J=10.8, 3.6 Hz, 1H), 3.44 (dd, J=11.2, 9.2Hz, 1H), 2.53-2.66 (m, 3H), 2.44 (dt, J=14.4, 3.6 Hz, 1H), 2.04 (m, 1H).

Intermediate 39

Step A

To a cooled (0° C.) solution of olefin INTERMEDIATE 38 (1.98 g, 14.1mmol) in THF (70 mL) was added dropwise 1.0 M BH₃.THF in THF (8.46 mL,8.46 mmol). The reaction mixture was warmed to room temperature andstirred for 1 h. Then a second portion of 1.0 M BH₃.THF in THF (8.46 mL,8.46 mmol) was added and the reaction mixture was stirred over theweekend. The reaction was quenched by the addition of water (70 mL),then was treated with NaBO₃.4H₂O (7.80 g, 50.8 mmol). The resultingsuspension was vigorously stirred for 4.25 h, then was concentrated todryness. The residue was purified by flash chromatography (silica, 5%methanol/DCM, then 8% methanol/DCM, then 10% methanol/DCM) to give 1.92g (85%) of diol as a mixture of isomers.Step B

Oxalyl chloride (110 □L, 1.26 mmol) was dissolved in DCM (8 mL),precooled to −78° C. Then a solution of DMSO (179 □L, 2.52 mmol) in DCM(1.5 mL) was added dropwise. After 5 min, a solution of the diol fromStep A immediately above (50.5 mg, 0.315 mmol) in DCM (1.5 mL) was addeddropwise. The reaction mixture was stirred at −78° C. for 20 min, thentriethylamine (702 □L, 5.04 mmol) was added dropwise. The reactionmixture was stirred for an additional 10 min, then was warmed to roomtemperature. After 45 min, the reaction mixture was poured into 2 N HClsolution and extracted three times with DCM. The combined organic layerswere washed with brine, dried over MgSO₄, filtered, and concentrated,giving 45.4 mg of crude product.

Intermediate 40

Olefin INTERMEDIATE 38 (2.00 g, 14.3 mmol) was dissolved in DCM (70 mL),cooled to −78° C., and treated with ozone gas (via pipet) until thereaction mixture appeared blue in color. Nitrogen gas was then bubbledthrough the reaction mixture until the blue color had disappeared(colorless). Triphenylphosphine (3.90 g, 14.9 mmol) was added and thereaction mixture was allowed to warm to room temperature and stir for1.25 h. The reaction mixture was concentrated and 50% ethylacetate/hexane was added to precipitate the triphenylphosphine oxide.The mixture was filtered and purified by MPLC (silica, ethyl acetate) togive 832 mg of the desired product. ¹H NMR (CDCl₃, 500 MHz): 9.78 (s,1H), 4.20-4.31 (m, 2H), 3.67 (dt, J=12.5, 3.0 Hz, 1H), 3.37 (t, J=11 Hz,1H), 3.21 (m, 1H), 2.89 (dd, J=18.5, 7.0 Hz, 1H), 2.72 (m, 1H), 2.40 (m,1H), 2.25 (dd, J=18.5, 5.5 Hz, 1H).

Intermediate 41

Step A

Through a cooled (−78° C.) solution of the allyl pyranone INTERMEDIATE38 (659 mg, 4.70 mmol) in methanol (10 mL) was bubbled ozone gas untilthe reaction appeared blue. Nitrogen gas was then bubbled through thesolution until the blue color disappeared. Sodium borohydride (267 mg,7.05 mmol) was added and the reaction mixture was permitted to warm tort. The reaction mixture was then concentrated and purified by flashchromatography (silica, 10% methanol/DCM) to afford 571 mg of diol as amixture of diastereomers.Step B

To a solution of the diol prepared as described in Step A (780 mg, 5.33mmol) and 2-nitrophenyl selenocyanate (1.33 g, 5.87 mmol) in THF (17 mL)at 0° C. was added dropwise tri-n-butylphosphine (1.59 mL, 6.40 mmol).The reaction mixture was warmed to rt, stirred for 45 min, andconcentrated. Purification by flash chromatography (silica, 80% ethylacetate/hexane) gave 1.30 g of hydroxyselenide intermediate (74%) whichwas combined with imidazole (671 mg, 9.85 mmol) in DMF (10 mL) andtreated with t-butyldimethylsilyl chloride (653 mg, 4.33 mmol) in DMF (5mL). The reaction mixture was stirred at room temperature for overnight.Although incomplete by TLC, the reaction was diluted with ether andwashed five times with water and once with brine. The ethereal phase wasdried over anhydrous MgSO₄, filtered, and concentrated. Purification byMPLC (silica, 40% ethyl acetate/hexane) provided 1.53 g of desiredproduct (87%).Step C

A cooled (0° C.) solution of the selenide prepared as described in StepB (1.50 g, 3.37 mmol) in THF (10 mL) was treated with 30% aqueous H₂O₂(2.92 mL, 33.7 mmol), allowed to warm to rt, and stirred for 2.5 h. Thereaction mixture was poured into 100 mL of 10% Na₂S₂O₃ solution, and theresulting mixture was extracted twice with ether. The combined ethereallayers were washed with saturated NaHCO₃ solution, then brined, driedover anhydrous MgSO₄, filtered, and concentrated. Purification by MPLC(silica, 20% ethyl acetate/hexane then 50% ethyl acetate/hexane)afforded 671 mg of olefin product as a mixture of isomers (˜4:1, 82%).¹H NMR (CDCl₃, 500 MHz, major isomer): 5.69 (m, 1H), 5.07-5.14 (m, 2H),3.95 (dt, J=11.5, 4.0 Hz, 1H), 3.89 (dd, J=4.0, 11.5 Hz, 1H), 3.57 (dt,J=4.5, 9.0 Hz, 1H), 3.44 (dt, J=2.5, 11.0H, 1H), 3.26 (dd, J=9.5, 11.5Hz, 1H), 2.24 (m, 1H), 1.84 (m, 1H), 1.61 (m, 1H), 0.90 (s, 9H), 0.066(s, 3H), 0.059 (s, 3H).Step D

Nitroso methylurea (4.42 g, 42.8 mmol) was added in portions to aprecooled (0° C.) two phase mixture of 40% KOH solution (18 mL) andether (52 mL). The mixture was swirled by hand until all of the solidshad dissolved and the ether layer had become deep yellow. This mixturewas cooled to −78° C. to freeze the aqueous layer, then the ethereallayer was decanted into a container with KOH pellets (˜5 g). Theresulting diazomethane solution was stored in the freezer for 0.5 h. Theolefin prepared as described in Step C (519 mg, 2.14 mmol) was dissolvedin ether (5 mL), cooled to 0° C., and combined with about ½ of thediazomethane solution. Then palladium acetate (12 mg) was added. Gasevolution and a color change from yellow to colorless was observed.After occasionally swirling this mixture for 15 min, MgSO₄ was added,and the mixture was filtered and concentrated to give 592 mg ofcyclopropyl product which required no further purification. ¹H NMR(CDCl₃, 500 MHz, major isomer): 3.91 (m, 2H), 3.70 (m, 1H), 3.48 (m,1H), 3.29 (dd, J=10.5, 14.5 Hz, 1H), 1.87 (m, 1H), 1.49-1.57 (m, 1H),0.92 (s, 9H), 0.72 (m, 1H), 0.54 (m, 2H), 0.39 (m, 1H), 0.31 (m, 1H),0.087 (s, 6H), 0.028 (m, 1H).Step E

A solution of the cyclopropyl-TBS-ether prepared as described in Step D(590 mg, 2.14 mmol) in THF (8 mL) was treated with 1.0 Mtetrabutylammonium fluoride solution in THF (2.57 mL, 2.57 mmol). Theresulting reaction mixture was stirred at room temperature for 24 h,then was concentrated and purified by MPLC (silica, 20% acetone/ether)to give 332 mg of product, which was contaminated by a small amount ofsolvent. ¹H NMR (CDCl₃, 500 MHz, major isomer): 3.98 (m, 1H), 3.93 (dd,J=4.5, 12.0 Hz, 1H), 3.68 (dt, J=4.5, 9.5 Hz, 1H), 3.44 (dt, J=2.5, 12.0Hz, 1H), 3.19 (dd, J=11.0, 11.5 Hz, 1H), 1.94 (m, 1H), 1.81 (m, 1H),1.58 (m, 1H), 0.77 (m, 1H), 0.62 (m, 1H), 0.44 (m, 1H), 0.37 (m, 1H),0.090 (m, 1H).Step F

To a cooled (−78° C.) solution of oxalyl chloride (0.373 mL, 4.28 mmol)in DCM (15 mL) was added dropwise DMSO (0.607 mL, 8.56 mmol) in DCM (2mL). After stirring at room temperature for 3 min, the alcohol preparedas described in Step E (2.1 mmol) in DCM (4 mL) was added dropwise.After an additional 15 min, triethylamine (2.39 mL, 17.1 mmol) was addeddropwise and the reaction mixture was permitted to warm to roomtemperature and stir for 1 h. The reaction mixture was diluted with DCMand washed in turn with 3N HCl solution and saturated NaHCO₃ solution,dried over anhydrous MgSO₄, filtered, and concentrated. Purification byMPLC (silica, ether) gave 272 mg of ketone product. ¹H NMR (CDCl₃, 500MHz): 4.10 (m, 2H), 3.87 (m, 1H), 3.68 (dd, J=8.0, 11.5 Hz, 1H),2.49-2.60 (m, 2H), 1.75 (m, 1H), 0.94 (m, 1H), 0.67 (m, 1H), 0.51 (m,1H), 0.22 (m, 1H), 0.14 (m, 1H).

Intermediate 42

Step A

This Intermediate was prepared following the procedures described inIntermediate 9 A-E, except that in Step D 2-iodopropane was used as thealkylating agent instead of acetone. Resolution of cis/transdiastereomers: Flash chromatography (silica gel, 8% EA/hexanes) H NMR(500 MHz, CDCl₃): 7.36 (m, 5H), 5.14 (s, 2H), 4.77 (m, 1H), 4.01 (d, J=

1H), 2.17 (m, 1H), 1.99-1.53 (m, 5H), 1.42 (m, 9H), 0.85 (d, J=7.0 Hz,6H).Step B

The BOC-amine from Step A (7.3 g, 20 mmol) was treated with hydrogenchloride (4N solution in dioxane). The reaction was allowed to stir for1.5 h at room temperature before being concentrated to remove thedioxane. The resultant solid was dissolved in DCM (150 mL) and treatedwith tetrahydro-4H-pyran-4-one (2.4 g, 24 mmol) and triethylamine (2.8mL, 20 mmol). The resulting solution was stirred at room temperature for5 minutes before 4 Å powdered molecular sieves (˜5 g) and sodiumtriacetoxyborohydride (17 g, 80 mmol) where added. The mixture wasstirred for 2 h at room temperature. The reaction was filtered throughcelite and washed with a saturated aqueous sodium bicarbonate solutionthen brine. The organic layer was dried over MgSO₄, filtered, andconcentrated under reduced pressure. To give 6.7 g of a colorless oil(97%). ESI-MS calc. for C21H31NO3: 345; Found: 346 (M+H).Step C

The amine from Step B (6.6 g, 19.1 mmol) was added to a solution of DCM(100 mL) and triethylamine (2.9 mL, 21 mmol). Trifluoroacetic anhydride(3.0 mL, 21 mmol) was added to the solution dropwise at room temperatureand the resulting solution was allowed to stir at room temperature for2.5 h. The reaction was diluted with DCM (100 mL) and washed withhydrochloric acid (1N aqueous solution) followed by brine. The organiclayer was dried over MgSO₄, filtered and concentrated under reducedpressure. The crude yellow oil was purified by MPLC (silica gel, 0 to30% EA/Hexanes) to give 4.9 g of a colorless oil (58%). ¹H NMR (CDCl₃,500 MHz): 7.37 (m, 5H), 5.18 (m, 2H), 4.20-3.88 (m, 4H), 3.64 (m, 1H),3.42 (t, J=12.0 Hz, 1H), 3.26 (t, J=11.5 Hz, 1H), 3.18 (t, J=11.5 Hz,1H), 2.81-2.65 (m, 2H), 2.26 (m, 1H), 1.89-1.80 (m, 3H), 1.64-1.40 (m,3H), 0.874 (m, 6H).Step D

The product from Step C (3.5 g, 7.9 mmol) was dissolved in methanol (60mL) and treated with 20% palladium hydroxide on activated carbon (350mg). This mixture was placed under a hydrogen atmosphere (1 atm) andallowed to stir at room temperature for 1.2 h. the reaction was filteredthrough celite and concentrated under reduced pressure to give 2.63 g ofa white solid (95%).

EXAMPLE 1

A solution of Intermediate 8 (50 mg, 0.13 mmol) and Intermediate 2 (18mg, 0.13 mmol) in DCM (5 mL) was treated with DIEA (35 μL, 0.19 mmol), 4Å powdered molecular sieves and sodium triacetoxyborohydride (108 mg,0.508 mmol) and stirred at room temperature overnight. The reaction wasquenched with saturated NaHCO₃ and extracted with DCM (5×). The combinedorganic phases were dried over Na₂SO₄ and concentrated. The crudeproduct was purified by preparative TLC (MeOH:DCM:NH₄OH/94.5:5:0.5) toyield the desired product (28 mg, 46%). Cis and trans pairs were furtherresolved by preparative TLC. The respective cis-racemate was furtherresolved by a chiral HPLC using a ChiralCel OD column. LC-MS forC₂₃H₃₁F₆N₂O₂ [M+H]⁺ calculated 481.22, found 481.30.

EXAMPLE 2

To a solution of the amine from Example 1 (cis-racemate, 25 mg, 0.052mmol), formaldehyde (37% aqueous solution, 12 μL, 0.16 mmol), DIEA (13μL, 0.078 mmol), TFA (5 μL) and MeOH (1.5 mL) was added NaCNBH₃ (17 mg,0.26 mmol). The reaction mixture was stirred at room temperatureovernight and concentrated in vacuo. Purification by preparative TLC(MeOH:DCM:NH₄OH/3:96.7:0.3) gave 17 mg (66%) of the desired product. Thecis-racemate was resolved by a ChiralPak AD semi-preparative HPLCcolumn. LC-MS for C₂₄H₃₂F₆N₂O₂ [M+H]⁺ calculated 495.24, found 495.25.

EXAMPLE 3

This amine was synthesized in 38% yield starting from the secondaryamine preparation of which was described in Example 1 according theprocedure described in Example 2, except that acetaldehyde was usedinstead of the formaldehyde. LC-MS for C₂₅H₃₄F₆N₂O₂ [M+H]⁺ calculated509.25, found 509.35.

EXAMPLE 4

This amine was prepared starting from the secondary amine preparation ofwhich was described in Example 1 according the procedure described inExample 2, except that propionaldehyde was used instead of theformaldehyde. LC-MS for C₂₆H₃₆F₆N₂O₂ [M+H]⁺ calculated 523.27, found523.20.

EXAMPLE 5

The amine from Example 1 (cis-racemate, 50 mg, 0.10 mmol) and methylbromoacetate (30 μL, 0.31 mmol) were dissolved in DCM (1 mL) and asaturated solution of NaHCO₃ (1 mL) was added. The reaction mixture wasstirred at room temperature overnight. Purified by preparative TLC(MeOH:DCM:NH₄OH (4:95.6:0.4) gave the desired product (43 mg, 74%).LC-MS for C₂₆H₃₄F₆N₂O₄ [M+H]⁺ calculated 553.24, found 553.25.

EXAMPLE 6

A solution of the amine from Example 2 (55 mg, 0.089 mmol), methyliodide (56 μL, 0.89 mmol), and THF (2 mL) was heated to 50° C. in asealed tube for 4.5 days and concentrated to dryness (74 mg, 99+%).L-C-MS for C₂₅H₃₅F₆N₂O₂ [M]⁺ calculated 509.26, found 509.35.

EXAMPLE 7

A mixture of the amine from Example 2 (50 mg, 0.089 mmol), H₂O₂ (30%aqueous solution, 3 mL), and MeOH (5 mL) was stirred at room temperaturefor a week before being concentrated to yield Example 7. L-C-MS forC₂₄H₃₂F₆N₂O₃ [M]⁺ calculated 510.23, found 511.3.

EXAMPLE 8

Step A

Intermediate 1 (2.00 g, 11.8 mmol), benzyl alcohol (6.22 mL, 57.6 mmol),DMAP (145 mg, 1.18 mmol), EDC (3.38 g, 17.6 mmol) and DCM (75 mL) weremixed together and stirred at room temperature. After completion ofreaction, the mixture was washed with H₂O (3×). The combined aqueouslayers were back extracted with DCM (1×). The combined organic layerswere washed with brine (1×), dried over MgSO₄, and concentrated todryness. The crude product was purified by flash chromatography(EtOAc:Hexanes/15:85) to yield the desired ester (900 mg, 29%). ¹H NMR(500 MHz, CDCl₃): 7.39-7.34 (m, 5H), 5.16 (d, J=2.8 Hz, 2H), 2.85 (d,J=18.3 Hz, 1H), 2.50-2.44 (m, 1H), 2.32-2.05 (m, 4H), 1.97-1.90 (m, 1H),0.94 (t, J=6.6 Hz, 6H).Step B

This compound was synthesized following the procedure described inExample 1. LC-MS for C₂₁H₃₂NO₃ [M+H]⁺ calculated 346.23, found 346.

EXAMPLE 9

Step A

Thionyl chloride (9.5 mL, 130 mmol) was added dropwise to methanol (225mL) and 3-fluorophenyl acetic acid (20 g, 130 mmol) was dumped into thesolution. The reaction mixture was refluxed for 1 h before beingconcentrated in vacuo to yield the desired product (23.4 g, >100%). ¹HNMR (400 MHz, CDCl₃): 7.30 (m, 1H), 7.02 (m, 3H), 3.73 (s, 3H), 3.64 (s,2H).Step B

The ester from Step A (23.25 g, 138.0 mmol) and 1,4-dichloro-cis-butene(15 mL, 0.14 mol) were dissolved in DME (200 mL) at 0° C. under nitrogenand NaH (60% dispersion in mineral oil, 14 g, 350 mmol) was added. Thereaction mixture was stirred for 12 h, quenched in ice water andextracted with ether (3×). The combined organic layers were washed withwater and brine, dried over anhydrous MgSO₄, and concentrated in vacuo.The crude product was purified by vacuum distillation (b.p.: 92-101° C.@ 0.11 mmHg) to yield the desired product (20 g, 60%), and about 20% ofthe corresponding cyclopropane product. ¹H NMR (500 MHz, CDCl₃): 7.29(m, 1H), 7.10 (m, 2H), 6.97 (m, 1H), 5.78 (s, 2H), 3.68 (s, 3H), 3.41(d, J=15.1 Hz, 2H), 2.78 (d, J=14.6 Hz, 2H).Step C

The olefin from Step B (12.5 g, 56.8 mmol), Borane (1 M in THF, 28.4 mL,28.4 mmol), and THF (100 mL) were mixed together and stirred at roomtemperature under nitrogen. Upon disappearance of the starting material,the reaction mixture was concentrated to dryness in vacuo and dissolvedagain in DCM. Anhydrous MgSO₄ (75 g) and PCC (49 g, 230 mmol) wereadded. The reaction mixture was stirred for 24 h and filtered throughsilica gel. The precipitate was suspended in DCM and ethyl acetate. Thesolution was refluxed for 20 minutes and filtered hot through silica gelto recover the product. The combined filtrate was concentrated in vacuoand purified by flash column chromatography (ethylacetate:hexanes/30:70) to yield the desired ketone (6.46 g, 48%). ¹H NMR(400 MHz, CDCl₃): 7.35 (m, 1H), 7.14-7.00 (m, 4H), 3.69 (s, 3H), 3.25(dd, J=17.9 Hz, 2.1 Hz, 1H), 2.98 (m, 1H), 2.61 (d, J=17.9 Hz, 1H),2.41-2.28 (m, 3H).Step D

Lithium hydroxide (2.05 g, 25.4 mmol) was dissolved in water (5 mL) anda solution of ester from Step C (3.0 g, 13 mmol) in methanol (25 mL) wasadded. The reaction mixture was stirred at room temperature for 5 hbefore being concentrated in vacuo. The concentrate was dissolved inwater and washed with ether. The aqueous layer was acidified to pH 2-3by addition of 2 N aqueous HCl and extracted with ether (4×). Thecombined organic layers were dried over anhydrous MgSO₄, filtered, andconcentrated in vacuo to yield the desired acid (2.678 g, 94%). Thecrude product was used in the next step without any additionalpurification.Step E

The above described acid (1.34 g, 6.57 mmol), 3-fluoro-5-trifluoromethylbenzylamine (972 μL, 6.57 mmol), HOAT (895 mg, 6.57 mmol) and EDC (1.9g, 9.85 mmol) were dissolved in DCM and stirred for 16 h at roomtemperature. The reaction mixture was diluted with methylene chloride,washed with 1 N HCl solution, saturated NaHCO₃, water, and brine. It wasdried over anhydrous MgSO₄ and concentrated in vacuo. The crude productwas purified by MPLC (ethyl acetate:hexanes/50:50) to yield the desiredamide (1.156 g, 44%). ¹H NMR (400 MHz, CDCl₃): 7.44 (m, 1H), 7.21 (d,J=8.0 Hz, 1H), 7.17 (m, 4H), 6.99 (d, J=9.0 Hz, 1H), 5.64 (s, 1H), 4.41(t, J=5.9 Hz, 2H), 3.21 (d, J=17.6 Hz, 1H), 2.80 (m, 1H), 2.64-2.44 (m,3H), 2.35 (m, 1H).Step F

The ketone from Step E (200 mg, 0.50 mmol), Intermediate 2 (70 mg, 0.50mmol), DIEA (130 μL, 0.76 mmol), sodium triacetoxyborohydride (534 mg,2.52 mmol), and 4 Å powdered molecular sieves were mixed together in DCMand stirred at room temperature for 24 h. The reaction was filteredthrough celite, concentrated, and purified by preparative TLC(methanol:NH₄OH:DCM (4:0.4:96.6) to yield the final product (156 mg,64%). LC-MS for C₂₅H₂₈F₅N₂O [M+H]⁺ calculated 483.20, found 483.25.

EXAMPLE 10

This compound was synthesized starting from the acid preparation whichwas described in Example 9, Step D and bis-trifluoromethylbenzylaminehydrochloride using the same procedure detailed in Example 9, Step E-F.LC-MS for C₂₆H₂₈F₇N₂O₂ [+H]⁺ calculated 533.20, found 533.2.

A number of additional compounds containing various aromatic groupsattached to Cl of the cyclopentane ring (R¹) in both the3,5-bistrifluoromethyl- as well as in the3-trifluoro-5-fluorobenzylamide series were prepared using proceduresanalogous to those described in Example 9. Table 1 summarizes thestructures and the calculated and observed MS characteristics of thesecompounds. TABLE 1

Molecular Calculated Found Example R₁ R₇ Formula [M + H⁺] [M + H⁺] 11

F C₂₆H₃₁F₄N₂O₃ 495.22 495.22 12

CF₃ C₂₇H₃₁F₆N₂O₃ 545.22 545.20 13

F C₂₃H₂₇F₄N₂O₂S 471.17 471.25 14

CF₃ C₂₇H₃₁F₆N₂O₂S 521.16 521.15 15

F C₂₃H₂₇F₄N₂O₂S 471.17 16

CF₃ C₂₄H₂₇F₆N₂O₂S 521.16 521.20 17

F C₂₅H₂₉F₆N₂O₂ 465.21 465.25 18

CF₃ C₂₆H₂₉F₆N₂O₂ 515.21 515.20Note:The respective cis- and trans- and trans diastereoisomeric pairs ofExample 12 were resolved by preparative TLC and the more polar cis-racemate was further resolved using chiral HPLC.

EXAMPLE 19

The amine from Example 1 (20 mg, 0.042 mmol), neat acetic anhydride (0.5mL), and pyridine (0.5 mL) were mixed and stirred at room temperature.Upon completion of the reaction, the mixture was concentrated, dissolvedin ether, washed with H₂O (3×), dried over MgSO₄, and concentrated todryness. The crude product was purified by preparative LC(EtOAc:Hexanes/50:50) to yield the desired product (5.5 mg, 25%). LC-MSfor C₂₅H₃₃F₆N₂O₃ [M+H]⁺ calculated 523.23, found 523.3.

EXAMPLE 20

Step A

A solution of Intermediate 9 (400 mg, 1.39 mmol), Intermediate 7 (470mg, 1.39 mmol), DIEA (365 μL, 2.09 mmol), and HOAT (190 mg, 1.39 mmol)in DCM (20 mL) was treated with EDC (400 mg, 2.09 mmol). The resultingmixture was stirred overnight, washed with saturated NaHCO₃, water (2×),brine, dried (anhydrous magnesium sulfate) and concentrated in vacuo.The crude product was purified by Preparative TLC (EtOAc:Hexanes/40:60)to yield the desired product (500 mg, 63%). ¹H NMR (500 MHz, CDCl₃):8.15 (bs, 1H), 7.84 (s, 1H), 7.53 (s, 1H), 5.26 (bs, 1H), 4.47 (d, J=6.1Hz, 2H), 4.01 (m, 1H), 2.60 (bs, 1H), 2.17 (dd, J=14.4 Hz, 5.4 Hz, 1H),2.07 (m, 1H), 2.00-1.87 (m, 3H), 1.61 (m, 1H), 1.43 (s, 9H), 1.27 (d,J=16.3 Hz, 6H). LC-MS for C₂₂H₃₁F₃N₂O₄[M+H]⁺ calculated 571.12, found571.2.Step B

The BOC-protected amine from the previous step (100 mg) was treated withHCl in dioxane (4 N, 6 mL) and was stirred at room temperature for 1 h.It was concentrated in vacuo to yield the desired amine hydrochloride(97 mg, 99%). LC-MS for C₁₇H₂₃F₃₁N₂O₂[M+H]⁺ calculated 470.07, found471.1.Step C

The amine from the previous step (400 mg, 0.851 mmol),tetrahydro-4H-pyran-4-one (170 μL, 1.70 mmol), and DIEA (225 μL, 1.28mmol) were dissolved in DCM (30 mL). Molecular sieves (4 Å, powdered)and sodium triacetoxyborohydride (900 mg, 4.26 mmol) were added. Thereaction mixture was stirred at room temperature overnight and thereaction was quenched with saturated NaHCO₃. The solution was heated at80° C. for 4 h to break up borane complexes, cooled to room temperatureand extracted with DCM (5×). The combined organic extracts were driedover Na₂SO₄ and concentrated to dryness. The crude product was purifiedby preparative TLC (MeOH:DCM:NH₄OH/5:94.5:0.5) to yield 270 mg (61%) ofthe desired product. L-C-MS for C₂₂H₃₁IF₃N₂O₃ [M+H]⁺ calculated 554.13,found 555.1.

Starting from Intermediates 9 and/or 10 a number of additional compoundswere synthesized. The applied procedures were analogous to thosedescribed in Example 20 varying the benzylamine in Step A as well as theketone in Step C. Their structure and MS characteristics are summarizedin Table 2. TABLE 2

Molecular Calc'd Found Ex. R1 R2 R3 R4 Formula [M⁺H⁺] [M⁺H⁺] 21

OH Cl CF₃ C₂₂H₃₁ClF₃N₂O₃ 463.19 463.15 22

OH H Ph C₂₇H₃₇N₂O₃ 437.27 437.35 23

OH H OCF₃ C₂₂H₃₂F₂N₂O₄ 445.22 445.3 24

OH H

C₂₂H₃₂F₃N₆O₃ 497.24 497.2 25

OH F CF₃ C₂₂H₃₁F₄N₂O₃ 447.22 445.25 26

OH Cl Cl C₂₁H₃₁Cl₂N₂O₃ 429.16 429.25 27

OH F CF₃ C₂₃H₃₃F₄N₂O₃ 461.23 461.25 28

OH F CF₃ C₂₉H_(34Cl)F₄N₂O₅ 601.20 601.3 29

OH F CF₃ C₂₂H₃₀F₅N₂O₃ 465.21 465.25 30

OH F CF₃ C₂₃H₃₀F₇N₂O₃ 515.21 515.2 31

OH F CF₃ C₂₃H₃₃F₄N₂O₄ 445.24 445.3 32

H CF₃ CF₃ C₂₇H₃₇F₆N₂O₃ 551.26 551.35 33

H CF₃ CF₃ C₂₄H₃₃F₆N₂O₂ 495.24 495.25 34

H CF₃ CF₃ C₂₅H₃₃F₆N₂O₃ 523.23 523.3 35

H CF₃ CF₃ C₂₇H₃₇F₆N₂O₃ 551.26 551.2 36

H CF₃ CF₃ C₂₄H₃₄F₆N₃O 494.25 494.3 37

H CF₃ CF₃ C₂₅H₃₄F₆N₃O₂ 522.25 522.25Notes:Example 27: The mixture of cis- and trans- racemates was separated bypreparative TLC (the less poiar racemate having the cis-relativestereochemistry).Example 30: The single enantiomers contained within the cis and transracemic pairs derived from the pyran ring were resolved using asemi-preparative ChiralCel OD column.Example 32: Cis and trans isomers on the cyclohexane ring were separatedby preparative TLC (MeOH : NH₄OH : DCM/1.5 : 0.15 : 98.35).

EXAMPLE 38

The ester described in Example 32 (25 mg, 0.043 mmol) was dissolved inMeOH (3 mL) and 5 N NaOH (1 mL) was added. After completion of reaction,the mixture was concentrated to dryness, the residue was dissolved inwater, acidified to pH 7, and extracted with DCM (6×). The combinedorganic phases were dried over Na₂SO₄ and concentrated in vacuo to yieldthe desired acid. It was further purified by reverse phase preparativeHPLC to afford 21.7 mg (91%) of the pure product. LC-MS for C₂₅H₃₃F₆N₂O₃[M+H⁺] calculated 523.23, found 523.2.

EXAMPLE 39

The compound in Example 39 was prepared as detailed in Example 38 usingthe ester described in Example 34 as starting material. LC-MS forC₂₄H₃₁IF₆N₂O₃ [M+H⁺] calculated 509.22, found 509.2.

EXAMPLE 40

The solution of the ester described in Example 35 (50 mg) in TFA (2.5mL) and DCM (2.5 mL) was stirred at room temperature. After completionof reaction, the reaction mixture was concentrated in vacuo and purifiedby reverse phase HPLC to yield the desired acid (0.64 mg). LC-MS forC₂₃H₂₉F₆N₂O₃ [M+H⁺] calculated 495.20, found 495.25.

EXAMPLE 41

Concentrated sulfuric acid (3 mL) was cooled to 0° C. before a solutionof the cis-racemate (50 mg) from Example 27 in acetonitrile (2 mL) wasadded. The mixture was stirred at room temperature overnight. Thereaction was poured onto ice, the solution was made basic with 5 N NaOH,and extracted with ether (3×). The combined organic phases were driedover anhydrous MgSO₄ and concentrated in vacuo. The crude product waspurified by reverse phase HPLC to yield the desired acetamide (1.7 mg).LC-MS for C₂₅H₃₆F₄₃O₃ [M+H⁺] calculated 502.26, found 502.3.

EXAMPLE 42

A solution of Intermediate 4 (110 mg, 0.698 mmol) in DCM (10 mL) wascooled to −78° C. and 03 was passed though until a permanent blue colorindicated completion of the reaction. The excess ozone was removed withnitrogen and the reaction mixture was warmed up to room temperature.This solution was dried over anhydrous MgSO₄ and after removal of thedrying agent by filtration, the corresponding amine Intermediate 14 (250mg, 0.558 mmol), DIEA (146 μL, 0.837 mmol), molecular sieves, and sodiumtriacetoxyborohydride (590 mg, 2.79 mmol) were added. The resultingmixture was stirred at room temperature overnight and filtered throughcelite. Saturated NaHCO₃ solution was added and the mixture was heatedto 60° C. for 3 h to break down the borane adducts. The product wasextracted with DCM (5×) and the combined organic layers were dried overNa₂SO₄, filtered, and concentrated in vacuo, and the product waspurified by preparative TLC (MeOH:DCM:NH₄OH/4:95.6:0.4) to yield thedesired product (23.3 mg). The two isomers were resolved by preparativeTLC. LC-MS for C₂₅H₃₃F₆N₂O₄ [M+H⁺] calculated 539.23, found 539.2.

EXAMPLE 43

This compound was prepared as described in Example 42. LC-MS forC₂₅H₃₃F₆N₂O₃ [M+H⁺] calculated 523.23, found 523.3.

EXAMPLE 44

The iodide described in Example 20 (100 mg, 0.18 mmol) and Pd(Ph₃P)₄ (22mg, 0.018 mmol) was dissolved in THF (10 mL) and Me₆Sn₂ (120 mg, 0.36mmol) was added. The reaction was complete after refluxing for 3 h. Themixture was allowed to cool to room temperature, diluted with EtOAc,washed with saturated NaHCO₃, dried over Na₂SO₄, and concentrated invacuo. The crude product was purified by preparative TLC(MeOH:DCM:NH₄OH/5:94.5:0.5) to yield the desired product (63.3 mg,59.4%). LC-MS for C₂₅H₄₀F₃N₂O₃Sn [M+H-1 calculated 591.19, found 593.2.

EXAMPLE 45

The tin-derivative described in Example 44 was stirred in 4 N HCl. LC-MSfor C₂₂H₃₂F₃N₂O₃ [M+H⁺] calculated 429.23, found 429.25.

EXAMPLE 46

The ester described in Example 28 (170 mg, 0.283 mmol) was dissolved inMeOH (2 mL) and sodium methoxide (0.5 M in MeOH, 640 μL, 0.3 mmol) wasadded. After completion of reaction, the mixture was concentrated invacuo and purified by preparative LC (MeOH:DCM:NH₄OH/6:93.4:0.6) toyield 79.7 mg (60%) of the desired product. LC-MS for C₂₂H₃₁F₄N₂O₄[M+H⁺] calculated 463.21, found 463.3.

EXAMPLE 47

This compound was prepared as detailed in Example 20, Step G startingwith the secondary amine from Example 27 and formaldehyde. LC-MS forC₂₄H₃₅F₄N₂O₃ [M+H⁺] calculated 475.25, found 475.3.

EXAMPLE 48

Step A

Iodine (2 crystals) was added to a suspension of magnesium (18.5 g, 761mmol) in anhydrous THF (240 mL) followed by cyclopropyl bromide (3 mL).The mixture was heated at 60° C. until the reaction initiated where uponcyclopropyl bromide (105 g, 868 mmol) was added at such a rate that agentle reflux was maintained. After complete addition the mixture washeated at reflux for an additional 1 h. To the cooled (ice-bath) mixturewas added tin (IV) chloride (15 mL, 130 mmol) and the reaction heated atreflux for 1 h. The reaction mixture was cooled and water (200 mL) wasadded cautiously, and the resulting mixture was extracted with Et₂O(3×400 mL). The combined Et₂O layers were dried over Na₂SO₄, filteredand concentrated in vacuo. The residue was purified by vacuumdistillation (b.p. 94° C. @ 1 mm Hg to give the desired product (19 g,52%).Step B

A mixture of the product from step A (4.0 g, 14 mmol), potassiumcarbonate (700 mg, 5 mmol), intermediate 20 (1.5 g, 5.0 mmol), andPd(PPh₃)₄ (300 mg, 0.25 mmol) in anhydrous N,N-dimethylformamide (50 mL)was heated at reflux for 1 h. The cooled reaction mixture was pouredinto water (350 mL) and extracted with EtOAc (3×200 mL). The combinedEtOAc layers were washed with water (3×300 mL), saturated NaCl (100 mL),dried over Na₂SO₄, filtered and evaporated. The residue was purified byMPLC (Biotage Flash 40) eluting with 2% EtOAc in hexanes to give thedesired product (650 mg, 62%); ¹H NMR 500 MHz (CDCl₃): 0.81 (2H, m),1.16 (2H, m), 2.02 (1H, m), 7.52 (1H, s), 7.54 (1H, s), 7.69 (1H,s).Step C

A solution of the product from step B (650 mg, 3.1 mmol) in a mixture ofethyl alcohol (25 mL) and ammonium hydroxide (5 mL) was hydrogenated at50 psi over Raney nickel (200 mg) for 7 h on a Parr apparatus. Thecatalyst was removed by filtration, and the filtrate evaporated. Theresidue was purified by MPLC (Biotage Flash 40) elution with 2% CH₃OH inCH₂Cl₂ containing 0.5% NH₄OH to give the desired product (336 mg, 51%);¹H NMR 500 MHz (CDCl₃): 0.74 (2H, m), 1.02 (2H, m), 1.95 (1H, m), 3.90(2H,s), 7.19 (1H, s), 7.22 (1H, s), 7.36 (1H,s).Step D

This compound was synthesized according to the procedure described inExample 146, except that Intermediate 9 was used instead of Intermediate10 and the previously described cyclopropylbenzylamine (Step C) insteadof the pyridylbenzylamine. The respective single diastereoisomers wereobtained by semi-preparative chiral HPLC using a ChiralCel OD column(eluent:hexane:ethanol/95:5, 9.0 mL/minutes. LC-MS for C₂₆H₃₇F₃N₂O₃[M+H]⁺ calculated 483.28, found 483.30.

EXAMPLE 49

Step A

This ester was synthesized as described in Intermediate 10, Step B.Step B

This intermediate was synthesized starting from the ester described inthe previous step according to the procedure detailed under thedescription of Intermediate 10, Step C, except that dimethyl disulfidewas used as alkylating agent instead of acetone. LC-MS for C₂₁H₂₄NO₂S[M+H⁺] calculated 354.14, found 354.25.Step C

The ester described in the previous step (2.10 g, 7.27 mmol) wasdissolved in MeOH (10 mL) and THF (10 mL) and a solution of lithiumhydroxide (1.5 g, 36 mmol) in H₂O (10 mL) was added. The mixture washeated to 60° C. overnight and concentrated in vacuo. The aqueous layerwas washed with hexanes, acidified to pH 4, and extracted with DCM (3×).The combined organic layers were dried over anhydrous MgSO₄ andconcentrated to dryness. The crude product was used in the followingstep without any additional purification.Step D

The amide group was attached in a procedure analogous to that describedin Example 20, Step A. LC-MS for C₂₁H₂₇N₂O₃S [M+H⁺] calculated 501.16,found 445.15 (loss of the t-butyl group).Step E

The BOC-protecting group was removed as described in Example 20, Step B.LC-MS for C₁₆H₁₈F₆N₂OS [M+H⁺] calculated 401.10, found 401.2.Step F

The final amine was prepared as detailed in Example 20, Step C. Therespective cis and trans isomers were resolved using preparative TLC(MeOH:DCM:NH₄OH/4:95.6: 0.4) with cis- being the desired isomer. LC-MSfor C₂₁H₂₇F₆N₂O₂S [M+H⁺] calculated 485.16, found 485.2.

EXAMPLE 50

Step A

The sulfide, preparation of which was described in Example 49, Step E(200 mg, 0.4 mmol) was dissolved in isopropanol (7 mL) before a solutionof oxone (500 mg, 0.8 mmol) in H₂O (7 mL) was added. The mixture wasstirred at room temperature for 2 h before being concentrated todryness. The concentrate was diluted with ether, washed with H₂O (3×),dried over anhydrous MgSO₄, and concentrated in vacuo to yield 207 mg(97%) of the desired compound. LC-MS for C₂₁H₂₇F₆N₂O₅S [M+H⁺] calculated533.15, found 433.15 (-BOC-group).Step B

This compound was prepared starting from the previously describedsulfone using the procedures detailed in Example 49, Steps E and F. Thecis- and trans-isomers were separated by preparative TLC with the lesspolar compound being the cis isomer. LC-MS for C₂₁H₂₇F₆N₂O₄S [M+H⁺]calculated 517.15, found 517.15.

EXAMPLE 51

This compound was prepared as detailed in Example 50 using Intermediate5 instead of tetrahydro-4H-pyranone. The two pairs of isomers wereseparated by preparative TLC (MeOH:DCM:NH₄OH/3:96.7:0.3). LC-MS forC₂₂H₂₉F₆N₂O₄S [M+H⁺] calculated 531.17, found 531.25.

EXAMPLE 52

A mixture of the product described in Example 51 (more polar isomer, 30mg, 0.058 mmol), formaldehyde (37% wt in H₂O, 15 μL, 0.17 mmol), TFA,NaCNBH₃ (20 mg, 0.29 mmol), and MeOH (5 mL) was stirred at roomtemperature overnight before being concentrated in vacuo and purified bypreparative TLC (MeOH:DCM:NH₄OH/4:95.6:0.4) to yield Example 52 (11 mg,35.7%). LC-MS for C₂₃H₃₁F₆N₂O₄S [M+H⁺] calculated 545.18, found 545.2.

EXAMPLE 53

The compound listed under Example 53 was prepared as detailed in Example49 except that diisopropyl sulfide was used instead of dimethyl sulfideand the cis and trans isomers were resolved in Step D instead of Step F.LC-MS for C₂₃H₃₁F₆N₂O₂S [M+H⁺] calculated 513 19, found 513.2.

EXAMPLE 54

The compound described under Example 54 was prepared as detailed inExample 50. LC-MS for C₂₃H₃₁F₆N₂O₄S [M+H]⁺ calculated 545.18, found545.2.

EXAMPLE 55

Step A

This compound was synthesized following the procedure detailed inExample 1.Step B

This compound was synthesized by reductive methylation of the secondaryamine from Step A following the procedure described in Example 52. LC-MSfor C₂₂H₃₅N₂O₂ [M+H⁺] calculated 359.26, found 359.35.Step C

Under N₂, the amide from the previous step (2.1 g, 5.9 mmol) wasdissolved in THF (20 mL) and a 1 M solution of borane in THF (29 mL, 29mmol) was added. The reaction mixture was refluxed overnight andconcentrated to dryness. The residue was dissolved in an 1% solution ofHCl in MeOH and heated to 50° C. overnight. The mixture was concentratedagain and dissolved in 1% HCl in MeOH solution to break down the excessborane. The crude product was used in the next step. LC-MS forC₂₂H₃₇N₂O₂ [M+H⁺] calculated 345.28, found 345.25.Step D

A mixture of the benzylamine from the previous step (2 g, 6 mmol),Pd(OH)₂ (500 mg), concentrated HCl (3 mL), and ethanol (50 mL) washydrogenated at 40 psi for 2 days on a Parr apparatus. The reactionmixture was filtered through celite and concentrated in vacuo to yield2.15 g, (99%) of the desired product.Step E

The final compound was prepared as detailed in Intermediate 8, exceptthat trifluoromethylbenzylamine was replaced by the amine from theprevious step and Intermediate 1 was replaced by benzoic acid. The crudeproduct was purified by preparative TLC (MeOH:DCM:NH₄OH/4:95.6:0.4).LC-MS for C₂₂H₃₅N₂O₂ [M+H⁺] calculated 359.26, found 359.35.

Following the procedure described in Example 55, by varying thestructure of the acids used in the last step of Example 55, a number ofanalogous amides were prepared. Their structure and MS characteristicsare summarized in Table 3. TABLE 3

Molecular Calculated Found Ex. R Formula [M⁺H⁺] [M⁺H⁺] 56

C₂₃H₃₃F₃N₂O₂ 427.25 427.3 57

C₂₄H₃₂F₆N₂O₂ 495.24 495.25 58

C₂₃H₃₃F₃N₂O₂ 427.25 427.3 59

C₂₃H₃₂F₄N₂O₂ 445.24 445.3 60

C₂₁H₃₃N₃O₃ 376.25 376.3 61

C₂₂H₃₃IN₂O₃ 501.15 501.25

EXAMPLE 62

A mixture of the amine described in Example 55, Step D (100 mg, 0.307mmol), DIEA (107 μL, 0.614 mmol), 5-bromoisatoic anhydride (75 mg, 0.31mmol), and DCM (20 mL) was stirred at room temperature overnight. Thefinal compound was obtained by preparative TLC(MeOH:DCM:NH₄OH/4:95.6:0.4) to yield 132 mg (82%) of the final product.LC-MS for C₂₂H₃₅BrN₃O₂ [M+H⁺] calculated 452.18, found 452.2.

EXAMPLE 63

A solution of the ketone Intermediate 12 (764 mg, 1.92 mmol), amineIntermediate 2 (264 mg of the hydrochloride, 1.92 mmol), 1.96 g of 4 Åpowdered molecular sieves and diisopropylethylamine (335 μL, 1.92 mmol)in dichloromethane (10 mL) was treated with sodium triacetoxyborohydride(2.03 g, 9.62 mmol) and the stirring at room temperature was continuedovernight. The reaction was quenched with a saturated aqueous solutionof sodium bicarbonate, and the crude product was extracted withdichloromethane. The combined organic extracts were washed with brineand dried with anhydrous sodium sulfate. The solvent was removed invacuo, and the residue (1.07 g) was purified by preparative TLC (sixplates, Silica gel, 1,000 micron, eluted with ethylacetate:ethanol:ammonium hydroxide/90:8:2) to yield 654 mg of homogenicproduct. The respective cis and trans racemates were separated usingpreparative TLC (eluent:dichloromethane:methanol:ammonium hydroxide(90:9:1). 317 mg (33%) of the higher eluting cis-racemate and 305 mg(32%) of the lower eluting trans racemic mixture were obtained. Thesingle enantiomers contained within the cis-racemic mixture wereobtained using a ChiralPak AD semi-preparative chiral HPLC column(eluent:hexane:ethanol/9:1, 9.0 mL/minute, Tr=6.99 min, and 11.12 min,respectively). cis-Isomer: ¹H NMR (500 MHz, CDCl₃): 10.0 (bs, 1H), 7.78(s, 1H), 7.73 (s, 2H), 4.53 (bd, J=15.10 Hz, 1H), 4.43 (bd, J=15.34 Hz,1H), 3.93 (bd, J=11.67 Hz, 1H), 3.82 (11.21 Hz, 1H), 3.57 (bs, 1H), 3.31(dt, J=11.89, 1.83 Hz, 1H), 3.25 (dt, J=12.13, 1.60 Hz, 1H), 2.65 (m,1H), 2.26 (m, 1H), 2.0 (m, 5H), 1.80 (bd, J=˜13 Hz), 1.28 (s, 3H), 1.20(m, 2H), 1.17 (s, 3H). LC-MS for C₂₃H₃₀F₆N₂O₃ [M+H]⁺ calc. 497.22, found497.35.

EXAMPLE 64

A solution of the alcohol described under Example 63 (cis-isomer, 80 mg,0.16 mmol) in 1.0 mL of acetonitrile was cooled to 0° C. andconcentrated sulfuric acid (380 mg, 3.88 mmol) were added. The solutionwas allowed to warm up to room temperature, and the reaction wascompleted by stirring at 50° C. for 1 h. Ice was then added, and theproduct was extracted into dichloromethane. After drying (anhydroussodium sulfate), the solvent was removed in vacuo, and the residue waspurified by preparative TLC (eluent:dichloromethane:methanol:ammoniumhydroxide/90:9:1) to get 29.8 mg (34%) of the pure product. LC-MS forC₂₅H₃₃F₆N₃O₃ [M+H]⁺ calc. 538.24, found 538.30.

EXAMPLE 65

Procedure A

A solution of the ketone Intermediate 12 (429 mg, 1.04 mmol), amineIntermediate 13 (as a hydrochloride salt, 158 mg, 1.04 mmol),diisopropylethylamine (182 μL, 1.04 mmol), 4 Å molecular sieves (766 mg)in 10 mL of dry dichloromethane was treated with sodiumtriacetoxyborohydride (1.10 g, 5.21 mmol) and the reaction mixture wasstirred at room temperature for 48 h. The crude reaction mixture waspoured onto a saturated solution of sodium bicarbonate (50 mL), and theorganic solvent was slowly evaporated under mildly reduced pressure (250torr) and slight heating (40° C.). HPLC analysis of the aqueous phaseindicated complete breakdown of the initial borane adduct (LC MS forC₂₄H₃₁BF₆N₂O₄ for [M+H]⁺ calc. 537.23, found 537.25) after approximately30 minutes. The crude product was extracted into dichloromethane, dried(anhydrous magnesium sulfate) and the solvent was removed in vacuo. Theresidue was further purified by preparative TLC to afford 243 mg (46%)of the higher eluting cis-isomeric mixture and 45.3 mg (8.5%) of thecorresponding lower eluting trans-isomeric pair. The cis-isomeric pairwas separated into single isomers by chiral semi-preparative HPLC usinga ChiralPak AD column, eluted by a mixture of hexanes and ethyl alcohol(95:5) at 9 mL/minutes. The retention time of the isomers under thecorresponding analytical conditions (1.0 mL/minutes flow rate) were 7.69and 12.33 minutes, for the biologically less and more active isomer,respectively. ¹H NMR (500 MHz, CDCl₃): 10.14 (s, 1H), 7.80 (s, 1H),7.73, s 2H), 5.80 (bs, 1H), 4.50 (bd, J=4.81 2H), 3.91 (bd, J=11.7 Hz,1H), 3.63 (bd, J=11.44 Hz, 1H), 3.56 (bs, 1H), 3.40 (dd, J=11.4, 2.3 Hz.1H), 3.33 (dt, J=11.45, 2.75 Hz, 1H), 2.72 (bs, 1H), 2.30 (m, 1H), 2.05to 1.85 (bm, 4H), 1.60 (m, 2H), 1.48 (m, 2H), 1.31 (s, 3H), 1.18 (s,3H), 0.88 (d, J=6.86 Hz, 3H). LC MS for C₂₄H₃₂F₆N₂O₃ for [M+H]⁺ calc.511.23, found 511.30.

Procedure B

A solution containing the ketone Intermediate 5 (667 mg, 5.84 mmol),Intermediate 14 (2.62 g, 5.84 mmol), diisopropylethylamine (1.02 mL,5.84 mmol) and crushed 4 Å molecular sieves (3.53 g) in dichloroethane(10 mL) was treated with sodium triacetoxyborohydride (6.18 g, 29.2mmol) and stirred at room temperature overnight. The workup wasidentical to that described under Procedure A of this example. The 2.39g of the product mixture was separated into single diastereomers using aChiralCel OD and ChiralPak AD columns. The obtained material (888 mg,30%) was identical in all respects to that obtained under Procedure A ofthis example.

EXAMPLE 66

A solution of the amine hydrochloride from Example 65 (slower elutingcis-isomer, in the form of a hydrochloride salt, 55 mg, 0.1 mmol),crushed 4 Å molecular sieves (226 mg) in dichloromethane (4 mL) wastreated with an aqueous solution of formaldehyde (210 μL, 37%, 2.0 mmol)followed by sodium triacetoxyborohydride (212 mg, 1.00 mmol) and themixture was stirred at room temperature overnight. It was poured ontosaturated aqueous sodium bicarbonate (20 mL), extracted withdichloromethane. The combined extracts were dried (anhydrous magnesiumsulfate), and the solvent was evaporated in vacuo. The residue waspurified by preparative TLC (eluent ethyl acetate:ethanol:ammoniumhydroxide/90:8:2) to afford 44 mg (87%) of the pure product. ¹H NMR (500MHz, CDCl₃): 9.18 (bs, 1H), 7.78 (s, 1H), 7.74 (s, 2H), 4.56 (dd,J=15.33, 6.17 Hz, 1H), 4.50 (dd, J=15.33, 5.49 Hz, 1H), 4.00 (dd,J=11.21, 4.58 Hz, 1H), 3.66 (d, J=11.44 Hz, 1H), 3.48 (dd, J=11.44, 2.06Hz, 1H), 3.30 (m, 2H), 2.75 (bd, J=12.12 Hz, 1H), 2.18 (s, 3H), 2.20 (m,3H), 1.80 (m, 5H), 1.37 (d, J=12.36 Hz, 1H), 1.29 (s, 3H), 1.21 (s, 3H),1.04 (d, J=7.09 Hz, 3H). LC MS for C₂₅H₃₄F₆N₂O₃ for [M+H]⁺ calc. 525.25,found 525.25.

EXAMPLE 67

This compound was synthesized starting from the secondary aminedescribed under Example 65 by a procedure analogous to that describedunder Example 66, except that formaldehyde was replaced by acetaldehyde.LC MS for C₂₆H₃₆F₆N₂O₃ for [M+H]⁺ calc. 539.26, found 539.25.

EXAMPLE 68

This compound was synthesized starting from amine Intermediate 14 andcyclohexanone in a procedure analogous to that described under Example65, Procedure B. LC MS: C₂₄H₃₂F₆N₂O₂ for [M+H]⁺ calc. 495.24, found495.20.

EXAMPLE 69

This compound was synthesized starting from amine Intermediate 14 and2-methylcyclohexanone in a procedure analogous to that described underExample 65, Procedure B. LC MS: C₂₅H₃₄F₆N₂O₂ for [M+H]⁺ calc. 509.25,found 509.40. Single diastereomers were obtained using a ChiralCel ODand ChiralPak AD semi-preparative columns.

EXAMPLE 70

This compound was synthesized starting from amine Intermediate 14 and3-cyclopentenone in a procedure analogous to that described underExample 65, Procedure B. LC MS: C₂₃H₃₀F₆N₂O₂ for [M+H]⁺ calc. 481.22,found 481.30.

EXAMPLE 71

This compound was synthesized starting from the secondary aminedescribed under Example 63 by a procedure analogous to that describedunder Example 66. LC MS: C₂₄H₃₂F₆N₂O₃ for [M+H]⁺ calc. 511.23, found511.23.

EXAMPLE 72

A solution of the ketone Intermediate 8 (983 mg, 2.48 mmol) and theamine Intermediate 15 (429 mg, 3.72 mmol) in neat Ti(OiPr)₄ (6 mL) wasstirred at room temperature overnight. Methanol (10 mL) was then addedvia syringe, followed by sodium borohydride (150 mg, 3.96 mmol) and thestirring at ambient temperature was continued for another 2 h. Thereaction mixture was then poured onto aqueous NaOH (0.1 N, 75 mL) andthe precipitate was filtered through a plug of Celite, which was thenwashed with methanol. The combined filtrates were concentrated in vacuoand the residue was extracted with ethyl acetate. The combined extractswere dried (anhydrous sodium sulfate) and the solvent was removed invacuo to leave 1.0092 g of crude product. Purification by preparativeTLC (six plates, 1000 micron, eluted withdichloromethane:methanol:ammonium hydroxide/92:8:1) afforded 574 mg(47%) of the higher eluting cis-racemic pair and 159 mg (13%) of thelower eluting trans-racemic product. LC MS: C₂₄H₃₂F₆N₂O₂ for [M+H]⁺calc. 495.24, found 495.35.

EXAMPLE 73

This compound was obtained starting from Intermediate 12 according to aprocedure described under Example 72. The single diastereomers containedwithin the cis-racemic pair were isolated using a ChiralPak ADsemi-preparative HPLC column. LC MS: C₂₄H₃₂F₆N₂O₃ for [M+H]⁺ calc.511.23, found 511.30.

EXAMPLE 74

This compound was obtained starting from the secondary amine describedunder Example 73 according to a procedure analogous to that describedunder Example 66. LC MS: C₂₅H₃₄F₆N₂O₃ for [M+H]⁺ calc. 525.25, found525.40.

EXAMPLE 75

Step A

A solution of methyl 3-methylene-1-isobutyl-cyclopentanecarboxylate (seeIntermediate 6, Step A, 3.92 g, 20.0 mmol) in a mixture of dioxane (50mL) and water (50 mL) containing 2.79 g (116 mmol) of lithium hydroxidemonohydrate was heated to reflux overnight. The solvent was removed invacuo, the residue was dissolved in water and the solution was madeacidic with 2 N HCl. The product was extracted from the aqueous phasewith chloroform (6×30 mL). The combined organic extracts were dried(anhydrous magnesium sulfate) and the solvent was removed in vacuo toyield 3.10 g (85%) of the desired product.Step B

A solution of the 3-methylene-1-isobutyl-cyclopentanecarboxylate (3.10g, 17.0 mmol) in dichloromethane was cooled to −78° C. and a stream ofozone was passed through the stirred solution until a permanent bluecolor indicated complete consumption of the olefin. The excess ozone waspurged with nitrogen, and triphenylphosphine (4.90 g, 18.7 mmol) wasadded. The cooling bath was removed, and the reaction mixture wasstirred at ambient temperature overnight. The solvent was evaporated invacuo, the residue was diluted with diethyl ether, and thetriphenylphosphine oxide was filtered off. The organic solution waswashed with aqueous 10% potassium carbonate (1×150 mL). The aqueousphase was washed with diethyl ether (3×50 mL), and made acidic with 2 NHCl. The desired acid was extracted into diethyl ether (4×50 mL), dried(magnesium sulfate) and the solvent was removed in vacuo to yield 2.72 g(87%) of the desired acid. ¹H NMR (CDCl₃, 500 MHz): 2.87 (dd, J=18.31,1.83 Hz, 1H), 2.43 (dp, J=6.64, 1.83 Hz), 2.30 (d, J=16.0 Hz, 1H), 2.30(d, 2.74 Hz, 1H), 2.15 (d, J=18.07 Hz, 1H), 1.94 (m, 2H), 1.70 (h,J=6.40 Hz, 1H), 1.57 (dd, J=13.96, 6.64 Hz, 1H), 0.93 (d, 6.63 Hz, 3H),0.92 (d, J=6.63 Hz, 1H).Step C

A mixture of the acid (750 mg, 4.07 mmol),3,5-bistrifluoromethylbenzylamine hydrochloride (1.138 g, 0.2180 mmol),diisopropylethylamine (710 μL, 4.07 mmol), 4-N,N-dimethylaminopyridine(60.0 mg 0.491 mmol) in dichloromethane (15 mL) was treated with1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EDC, 1.56g, 8.14 mmol) and stirred at room temperature for 24 h. The reactionmixture was diluted with dichloromethane (20 mL), washed with water(3×30 mL), brine (1×30 mL), dried over anhydrous sodium sulfate and thesolvent was evaporated under reduced pressure to yield 1.25 g (75%) ofthe desired product which was further purified by column chromatography(silica gel, ethyl acetate:hexanes (1:1) to yield 583 mg (35%) of thepure desired product. LC-MS for C₁₉H₂₂F₆NO₂ [M+H]⁺ calculated 410.15,found 410.20.Step D

The final amine was synthesized starting from Intermediate 2 and theketone described in the previous step according to the proceduredescribed under Example 1. LC-MS for C₂₄H₃₂F₆N₂O₂ [M+H]⁺ calculated495.24, found 495.30.

EXAMPLE 76

Step A

This acid was synthesized in a procedure analogous to that described forthe preparation of Intermediate 11, except that the acetone in Step Cwas replaced by N-Benzyloxycarbonylpiperidin-4-one. LC-MS for C₁₉H₂₃NO₆[M+H]⁺ calculated 362.15, found 362.15.Step B

This keto amide was synthesized starting from the previously describedacid and 3,5-bistrifluoromethyl-benzylamine according to the proceduredescribed for the preparation of Intermediate 12. ¹H NMR (CDCl₃, 500MHz): 7.78 (m, 2H), 7.72 (s, 2H), 7.35 (bm, 4H), 5.12 (s, 2H), 4.58 (dd,J=15.6, 6.0 Hz, 1H), 4.52 (dd, J=15.3, 5.7 Hz, 1H), 4.08 (bs, 2H), 3.05(s, 2H), 2.74 (d, J=18 Hz, 1H), 2.40 to 2.15, bm, 5H), 1.9 to 1.4 (bm,6H). LC-MS for C₂₈H₂₈N₂F₆O₅ [M+H]⁺ calculated 587.19, found 587.35.Step C

The final amine was synthesized starting from the Intermediate ketonedescribed in the previous step and Intermediate 2 according to aprocedure analogous to that described for the preparation of Example 63.The respective single diastereomers were obtained by a chiral HPLCseparation using a ChiralPak AD semi-preparative column,eluent:hexane:ethanol/80: 20, flow rate of 9.0 ml/minutes. LC-MS forC₃₃H₃₉F₆N₃O₅ [M+H]⁺ calculated 672.28, found 672.30.

EXAMPLE 77

A solution of the CBZ-Protected amine, the synthesis of which wasdescribed under Example 76 (109 mg, 0.154 mmol), was dissolved inethanol (15 mL), and Pd/C (10%, 27 mg) was added, and the resultingmixture was hydrogenated under a hydrogen filled balloon at roomtemperature for 24 h. The catalyst was filtered off, and the solvent wasremoved in vacuo to yield 87.7 mg (93%) of the desired product. LC-MSfor C₃₅H₃₃F₆N₃O₃ [M+H]⁺ calculated 538.24, found 538.30.

EXAMPLE 78

The compound described under this example was synthesized using aprocedure analogous to that described for preparation of Example 76,except that in Step A the N-CBZ-piperidin-4-one was replaced bytetrahydro-4H-pyran-4-one. The respective single diastereomers wereobtained by a chiral HPLC separation using a ChiralPak ADsemi-preparative column, eluent:hexane:ethanol/90:10, flow rate of 9.0mL/minutes. LC-MS for C₂₅H₃₂F₆N₂O₄ [M+H]⁺ calculated 539.23, found539.35.

EXAMPLE 79

The compound described under this example was synthesized using aprocedure analogous to that described for preparation of Example 76,except that in Step A the N-CBZ-piperidin-4-one was replaced bydicyclopropylketone. LC-MS for C₂₇H₃₄F₆N₂O₃ [M+H]⁺ calculated 549.25,found 549.40.

EXAMPLE 80

Step A

A solution of 3-oxocyclopentanecarboxylic acid (6.20 g, 48.4, Stetter,H., Kuhlman, H., Liebigs Ann. Chemie, 1979, 7, 944-9) and3-methyl-2-buten-1-ol (5.90 mL, 58.1 mmol) and DMAP (140 mg) indichloromethane (50 mL) was treated with EDC and stirred at roomtemperature overnight. The reaction was quenched by pouring onto 100 mLof water, and the product was extracted with dichloromethane. Thecombined organic extracts were washed with brine and dried withanhydrous magnesium sulfate. Evaporation of the solvent in vacuo gave10.51 g (100%) of the desired ester. ¹H NMR (CDCl₃, 500 MHz): 5.33 (bt,J=7.33 Hz, 1H), 4.62 (d, J=7.32 Hz, 2H), 3.12 (ddd, J=14.9, 8.0, 6.9 Hz,1H), 2.55 to 2.10 (m, 6H0, 1.77 (s, 3H), 1.72 (s, 3H).Step B

A solution of the ester from the previous step (10.50 g, 53.78 mmol) andTsOH (500 mg) in dichloromethane (50 mL) was treated with trimethylorthoformate (24 mL, 220 mmol) and stirred at room temperatureovernight. The reaction was quenched by pouring onto a saturatedsolution of sodium bicarbonate and the crude product was extracted withdichloromethane. The combined organic layers were washed with brine,dried with anhydrous magnesium sulfate, and the solvent was evaporatedin vacuo. The residue was further purified by distillation (b.p.: 123°C. @ 4 mm Hg) to give 8.27 g (63%) of the desired acetal. ¹H NMR (CDCl₃,500 MHz): 5.33 (bt, J=7.09 Hz, 1H), 4.57 (d, J=7.32 Hz, 2H), 3.21 (s,3H), 3.19 (s, 3H), 2.87 m, 1H), 2.15 to 1.80 (bm, 6H), 1.76 (s, 3H),1.70 (s, 3H).Step C

A solution of diisopropylamine (14.28 mL, 101.9 mmol) in THF (200 mL)was cooled to −78° C. and n-butyllithium (40.76 mL, 2.5 M in hexane, 110mmol) was added via syringe. After 10 minutes, the neat ester from theprevious step (12.34 g, 50.94 mmol) was added via syringe, followedafter 20 minutes by neat chloro trimethylsilane (12.93 mL, 101.9 mmol).The solution was allowed to warm up to room temperature over 3 h, afterwhich time it was quenched by pouring onto a 10% aqueous solution ofcitric acid. The crude product (19.75 g) was obtained by extraction withdiethyl ether, drying (magnesium sulfate) and evaporation of the solventin vacuo. It was further purified by flash chromatography (silica gel,ethyl acetate:hexane/2:3) to yield 5.25 g (43%) of the pure product. ¹HNMR (CDCl₃, 500 MHz): 5.95 (dd, J=17.2, 10.8 Hz, 1H), 5.07 (dd, J=10.75,0.9 Hz, 1H), 5.25 (dd, J=17.40, 0.9 Hz, 1H), 3.21 (s, 3H), 3.14 (s, 3H),2.58 (d, J=13.73 Hz, 1H), 2.28 (m, 1H), 1.88 to 1.76 (bm, 3H), 1.73 to1.66 (bm, 2H), 1.084 (s, 3H), 1.00 (s, 3H). ¹³C NMR (CDCl₃, 125 MHz):174.7, 143.9, 113.1, 112.3, 58.7, 53.8, 50.7, 41.4, 36.5, 31.4, 24.7,23.1, 22.0.Step D

A solution of the crude acid, the preparation of which was described inthe previous step (1.0 g, 4.2 mmol). 3,5-bistrifluoromethylbenzyl aminehydrochloride (1.15 g, 4.13 mmol), diisopropylethylamine (0.72 mL, 4.13mmol) in dichloromethane (12 mL) was treated with EDC (1.18 g, 6.19mmol) and stirred at room temperature overnight. The reaction mixturewas diluted with dichloromethane, washed with water and brine, driedwith magnesium sulfate and the solvent was evaporated in vacuo to leave2.00 g of crude product. Flash chromatography (silica gel, ethylacetate:hexanes/1:1) gave 1.38 g (72%) of the pure amide. ¹H NMR (CDCl₃,500 MHz): 7.78 (s, 3H), 6.30 (bt, J=5.72 Hz, 1H), 17.4, 10.8 Hz, 1H),5.06 (dd, J=10.8, 1.1 Hz, 1H), 5.00 (dd, J=17.4, 0.9 Hz, 1H), 4.57 (dd,J=15.6, 6.2 Hz, 1H), 4.53 (dd, J=15.3, 6.0 Hz, 1H), 3.20 (s, 3H), 3.08(s, 3H), 2.47 (d, J=14.2 Hz, 1H), 2.14 (m, 1H), 1.95 to 1.70 (bm, 4H),1.07 (s, 3H), 1.06 (s, 3H).Step E

A solution of the acetal from previous step (407 mg, 0.871 mmol) wasdissolved in dichloromethane (6.0 mL) and treated with TFA (1.0 mL). Thesolution was stirred at room temperature overnight. The reaction mixturewas poured onto aqueous saturated sodium bicarbonate and extracted withdichloromethane. Drying (magnesium sulfate) and evaporation of thesolvent in vacuo gave 326 mg (89%) of the pure ketone. ¹H NMR (CDCl₃,500 MHz): 7.79 (s, 1H), 7.72 (s, 2H), 6.58 (t, J=5.26 Hz, 1H), 6.02 (dd,J=17.40, 10.75 Hz, 1H), 5.16 (d, J=10.76 Hz, 1H), 5.08 (d, J=17.60 Hz,1H), 4.52 (d, J=5.95 Hz, 2H), 2.78 (dd, J=18.08, 1.37 Hz, 1H), 2.35 (m,3H), 2.12 (m, 2H), 1.09 (s, 6H). ¹³C NMR (CDCl₃, 125 MHz): 216.2, 174.4,144.7, 141.3, 128.1, 121.7, 114.9, 58.9, 44.5, 43.4, 40.6, 37.0, 28.7,24.3, 23.5. LC-MS for C₂₀H₂₁F₆NO₂ [M+H]⁺ calculated 422.15, found422.20.Step F

The final amine listed under this example was synthesized starting fromthe ketone from the previous step and Intermediate 2 in a procedureanalogous to that described under Example 1. LC-MS for C₂₅H₃₂F₆N₂O₂[M+H]⁺ calculated 507.24, found 507.40.

EXAMPLE 81

A solution of the olefin, the preparation of which was described underExample 80 (89 mg, 0.16 mmol), in ethanol (20 mL) was treated withperchloric acid (500 μL), cooled to −78° C. and ozone was passed throughuntil a permanent blue color indicated completion of the reaction. Theexcess ozone was purged with a stream of nitrogen, and sodiumborohydride (150 mg) was added. The temperature was allowed to rise toroom temperature overnight, another 400 mg of sodium borohydride wasadded, and the reaction was stirred at ambient temperature another 2 h.The solvent was removed in vacuo, and the residue was treated with anaqueous saturated solution of sodium bicarbonate. The crude product wasextracted into dichloromethane, dried with anhydrous sodium sulfate, andthe solvent was removed in vacuo. The respective cis- (higher eluting,18.7 mg, 22%) and trans- (lower eluting, 10.0 mg, 12%) racemic pairswere obtained by preparative TLC (dichloromethane:methanol:ammoniumhydroxide/90:9:1). LC-MS for C₂₄H₃₂F₆N₂O₃ [M+H]⁺ calculated 511.23,found 511.40. The respective single diastereomers contained within thecis-pair were separated by semi-preparative HPLC (ChiralPak AD, 93%hexane:7% ethanol, 9 mL/min). The retention times of an analogousanalytical run (1.0 mL/min) were Tr=7.65 and 9.98 minutes.

EXAMPLE 82

A solution of the olefin, the preparation of which was described underExample 80 (82 mg, 0.151 mmol), and Pd/C (50 mg, 10%) in ethanol (10 mL)was placed under a hydrogen balloon at room temperature for 30 minutes.The catalyst was filtered off, and the solvent was removed in vacuo toyield 61.8 mg (75%) of the pure product. LC-MS for C₂₅H₃₄F₆N₂O₂ [M+H]⁺calculated 509.25, found 509.35.

EXAMPLE 83

Step A

A solution of diisopropylamine (2.71 mL, 19.3 mmol) in diethyl ether (60mL) was cooled to −78° C. and n-butyllithium (7.73 mL, 19 mmol, 2.5 Msolution in hexane) was added drop-wise, via syringe. A solution of theallyl ester (5.86 g, 17.58 mmol, prepared from(3R,1S)-3-benzhydrylimonocyclopent-4-ene-carboxylic acid and allylbromide as described under Preparation of Intermediate 9, Step A) indiethyl ether (30 mL) was then added and the solution was stirred at−78° C. 2.5 h. At this point a solution of zinc chloride in diethylether (19.33 mL, 1.0 M, 19 mmol) was added dropwise, followed by 2.58 mL(35.1 mmol) of neat acetone. The reaction mixture was stirred at −78° C.for another 45 minutes, after which it was quenched by the addition of300 mL of an aqueous saturated solution of ammonium chloride. Theaqueous layer was separated and washed with ether three more times. Thecombined organic extracts were dried with anhydrous sodium sulfate andthe solvent was evaporated in vacuo. The obtained mixture of the cis-and trans-diastereoisomer (6.35 g) was used without purification in thenext step.Step B

The crude Schiff base from the previous step (6.35 g) was dissolved inTHF (30 mL) and 2 N aqueous HCl (10 mL) was added and the mixture wasstirred at room temperature overnight. The solvent was evaporated todryness under reduced pressure, and the obtained mixture of the desiredamines and benzophenone (7.51 g) was used in the next step without anyfurther purification.Step C

A solution of the crude amine from the previous step (7.51 g) indichloromethane (50 mL) was treated with di-tert-butyl dicarbonate (5.35g, 24.5 mmol) and a saturated aqueous solution of sodium bicarbonate (50mL) was added. The biphasic reaction mixture was vigorously stirred for1 hr, and the organic layer was separated. The aqueous phase wasextracted two more times with dichloromethane, and the combined organicextracts were dried, and the solvent was removed in vacuo. The residue(9.44 g) was further purified by gradient column chromatography(eluent:ethyl acetate:hexanes, 15% to 100%). 744 mg (14%, three steps)of the first eluting 1,3-trans-isomer, 405 mg (8%) of the second eluting1,3-cis-isomer and 658 mg of mixed fractions where obtained in anoverall yield of 34% for three steps.

Higher Eluting 1,3-trans-Isomer: ¹H NMR (CDCl₃, 500 MHz): 5.93 (m, 3H),5.37 (dd, J=17.2, 1.2 Hz, 1H), 5.30 (dd, J=10.3, 0.7 Hz, 1H), 4.70 (m,3H), 2.63 (dd, J=14.5, 8.2 Hz, 1H), 1.95 (dd, J=14.7, 4.6 Hz, 1H), 1.45(s, 9H), 1.17 (s, 6H).

Lower Eluting 1,3-trans-Isomer: ¹H NMR (CDCl₃, 500 MHz): 5.90 (m, 3H),5.35 (dd, J=17.2, 1.1 Hz, 1H), 5.25 (dd, J=10.5, 0.7 Hz, 1H), 4.85 (bs,1H), 0.72 (bs, 1H), 4.60 (d, J=5.72 Hz, 2H), 2.80 (dd, J=14.0, 8.24 Hz,1H), 1.77 (dd, J=14.4, 5.5 Hz, 2H), 1.46 (s, 9H), 1.23 (s, 3H), 1.22 (s,3H).Step D

A solution of the ester from previous step (lower eluting cis-isomer,640 mg, 1.96 mmol) and morpholine (1.71 mL, 19.6 mmol) in THF (20 mL),was thoroughly degassed (vacuum/nitrogen cycle) and Pd(Ph₃P)₄ (237 mg)was added. The reaction mixture was stirred at room temperature for 2 h,after which time it was diluted with dichloromethane and washed with 2 Naqueous HCl. The organic phases were dried (magnesium sulfate) and thesolvent was removed in vacuo to yield 685 mg of crude product. It wasused in the next step without further purification.Step E

A solution of the crude acid from the previous step (685 mg, max 1.96mmol), 3,5-bistrifluoromethylbenzylamine hydrochloride (1.64 g, 5.88mmol), diisopropylethylamine (993 μL, 5.88 mmol), HOAT (800 mg, 5.88mmol) in dichloromethane (40 mL) was treated with EDC (1.13 g, 5.88mmol) and the reaction mixture was stirred at room temperatureovernight. It was diluted with dichloromethane and washed with water.After drying (anhydrous sodium sulfate) the solvent was evaporated todryness. The residue was further purified by column chromatography(silica gel, ethyl acetate:hexane/2:3) to yield 555.6 mg (54%, twosteps) of the pure amide. ¹H NMR (CDCl₃, 500 MHz): 7.85 (bs, 1H), 7.78(s, 1H), 7.75 (s, 2H), 5.98 (dd, J=5.5, 2.1 Hz, 1H), 5.84 (dd, J=5.50,1.83 Hz, 1H), 4.98 (bs, 1H), 4.55 (m, 3H), 4.2 (bs, 1H), 2.75 (dd,J=14.9, 9.2 Hz, 1H), 2.02 (dd, J=14.6, 4.6 Hz, 1H), 1.38 (s, 9H), 1.28(s, 3H), 1.15 (s, 3H).Step F

The BOC-Protected amine from the previous step (516 mg, 1.01 mmol) wasdissolved in 8 mL of 4 N HCl solution in dioxane. After 30 minutes ofstirring at room temperature the solvent was removed in vacuo, and thecrude hydrochloride (454 mg, 100%) was used in the next step withoutfurther purification.Step G

The final product was obtained from the previously described amine andtetrahydro-4H-pyran one as described in the procedure from Example 65,Procedure B. ¹H NMR (CDCl₃, 500 MHz): 9.82 (bs, 1H), 7.79 (s, 1H), 7.68(s, 2H), 6.24 (d, J=5.72 Hz, 1H), 6.03 (bs, 1-H), 5.94 (dd, J=5.72, 2.52Hz, 1H), 4.58 (dd, J=15.6, 6.2 Hz, 1H), 4.40 (dd, J=15.6, 5.04 Hz, 1H),4.16 (bd, J=7.32 Hz, 1H), 3.90 (bd, J=11.7 Hz, 2H), 3.30 (m, 2H), 2.67(m, 1H), 2.20 (dd, J=14.41, 7.3 Hz, 1H), 1.95 (d, J=14.42 Hz, 1H), 1.70(bd, J=12.8 Hz, 1H), 1.63 (bd, J=13.0 Hz, 1H), 1.30 (s, 3H), 1.25 (bm,2H), 1.12 (s, 3H). LC-MS for C₂₃H₂₉F₆N₂O₃ [M+H]⁺ calculated: 495.20,found 495.25.

EXAMPLE 84

This compound was synthesized starting from the trans-isomeric esterintermediate, described under Step C, Example 83, following theprocedures described under Example 83, Steps D-G. LC-MS for C₂₃H₂₈F₆N₂O₃[M+H]⁺ calculated: 495.20, found 495.30.

EXAMPLE 85

This compound was prepared following the procedure described for thepreparation of the compound under Example 83, except that in Step Gtetrahydro-4H-pyran-4-one was replaced by Intermediate 5. The respectivecis- and trans-diastereoisomeric pairs (THP ring) were separated usingpreparative TLC (ethyl acetate:ethanol:ammonium hydroxide/85: 4:1) andthe respective isomers contained within the higher eluting cis-pair wereobtained by separation on a semi-preparative chiral HPLC column:ChiralPak AD, 95% hexanes, 9.0 ml/minutes. The respective retentiontimes of an analogous analytical run (1.0 mL/min) were 11.8 and 15.0minutes, respectively. LC-MS for C₂₃H₂₈F₆N₂O₃ [M+H]⁺ calculated: 495.20,found 495.30.

The slower (Tr=15.0 minutes) eluting cis-diastereoisomer: ¹H NMR (CDCl₃,500 MHz): 9.9 (bs, 1H), 7.78 (s, 1H), 7.67 (s, 2H), 6.27 (m, 1H), 5.95(m, 1H), 4.60 (m, 1H), 4.40 (m, 1H), 4.10 (d, J=18.1 Hz, 1H), 3.85 (bt,J=11.9 Hz, 1H), 3.65 (bt, J=13.0, 2.8 Hz, 2H), 3.42 (bd, J=11.44 Hz,1H), 3.32 (m, 1H), 2.82 (m, 1H), 2.20 (m, 1H), 1.98 (m, 1H), 1.76 (bs,1H), 1.40 (m, 1H), 1.30 (s, 3H), 1.10 (s, 3H), 0.82 (d, J=7.10 Hz, 3H).

EXAMPLE 86

This compound was prepared from the amine, the synthesis of which wasdescribed under Example 86 and formaldehyde following the proceduredescribed in Example 66. ¹H NMR (CDCl₃, 500 MHz): 7.80 (s, 1H), 7.71 (s,2H), 6.05 (dd, J=5.72, 2.29 Hz, 1H), 5.92 (bd, J=4.81 Hz, 1H), 5.30 (bs,1H), 4.56 (d, J=5.95 Hz, 1H), 4.2 (bs, 1H), 4.0 (dd, J=12, 4.5 Hz, 1H),3.73 (d, J=11.44 Hz, 1H), 3.47 (dd, J=11.44, 1.83 Hz, 1H), 3.32 (dt,J=12.1, 2.3 Hz, 1H), 2.57 (dt, J=11.9, 4.1 Hz, 1H), 2.32 (dd, J=14.6,8.2 Hz, 1H), 2.02 (s, 3H), 1.97 (dd, J=14.9, 4.6 Hz, 1H), 1.85, (bs,1H), 1.75 (m, 2H), 1.52 (bd, J=11.44 Hz, 1H), 1.21 (s, 3H), 1.14 (s,3H), 1.02 (d, J=6.86 Hz, 3H). LC-MS for C₂₅H₃F₆N₂O₃ [M+H]⁺ calculated:523.23, found 523.30.

EXAMPLE 87

Step A

A solution of diisopropylamine (2.70 mL, 19.3 mmol) in tetrahydrofuran(20 mL) was cooled to −78° C. and a solution of n-butyllithium inhexanes (7.70 mL, 2.5M, 19.3 mmol) was added via syringe, followed by asolution of the Schiff base, the preparation of which was described inIntermediate 9, Steps A to C (5.685 g, 14.82 mmol), in THF (10 mL). Theenolate was allowed to form for 3 h at −78° C., after which time theneat acetaldehyde (1.00 mL, 29.7 mmol) was added. The reaction wasquenched with the addition of aqueous citric acid (200 mL, 10%) and thecrude product was extracted into diethyl ether. Drying (anhydrousmagnesium sulfate) and evaporation of the solvent gave the crude desiredproduct (6.16 g). This was further purified by flash chromatography(deactivated silica gel, ethyl acetate:hexanes/3:7) to yield the desiredcis-isomer (2.32 g, 54%). This Schiff base was found to be unstable, andwas used in the next step without delay. LC-MS for C₂₉H₂₉NO₃ [M+H]⁺calculated: 428.21, found 428.20.Step B

The Schiff base prepared in the previous step (2.323 g, 5.433 mmol) wasdissolved in THF (20 mL) and 2 N aqueous HCl was added. The reactionmixture was stirred at room temperature 2 h, after which time thevolatiles were removed in vacuo. The resulting mixture of the desiredamine hydrochloride and benzophenone was used in the next step withoutfurther purification.Step C

The crude product from the previous step (max 5.433 mmol) was dissolvedin dichloromethane (50 mL) and di-tert-butyl dicarbonate (2.371 g, 10.86mmol) was added followed by 50 mL of a saturated solution of sodiumbicarbonate. The reaction mixture was vigorously stirred at roottemperature for 1 h. The layers were separated and the aqueous phase waswashed with dichloromethane. The combined organic extracts were dried(anhydrous magnesium sulfate) and the solvent was evaporated in vacuo.Final purification by gradient flash chromatography (ethylacetate:hexanes/0% to 40%) gave the desired BOC-protected amine (619 mg,32%, two steps) as a diastereoisomeric mixture (3:2) of two isomers.LC-MS for C₂₀H₂₉NO₅ [M+H]⁺ calculated: 364.20, found 264.20 (loss of theBOC group).Step D

This acid was prepared following the procedure described in Intermediate9, Step F, and was used in the next step without further purification.Step E

The solution of the acid from the previous step (63 mg, 0.23 mmol) and3,5-bistrifluoromethylbenzyl amine hydrochloride (77 mg, 0.28 mmol),1-hydroxy-7-azobenzotriazole (31.5 mg, 0.231 mmol) in dichloromethane (6mL) was treated with EDC (66 mg, 0.35 mmol) and the resulting mixturewas stirred at room temperature overnight. The reaction was quenchedwith water, and the product was extracted into dichloromethane. Thecombined organic extracts were dried (anhydrous magnesium sulfate) andthe solvent was removed in vacuo. The residue (112.7 mg) was furtherpurified by preparative TLC (ethyl acetate:hexanes/3:2) to yield thedesired product (41 mg, 36%) as a mixture of two diastereoisomers. Thesewere separated by preparative TLC (DCM:acetone/9:1) to yield singleisomers (the hydroxyethyl side-chain) of unknown absolutestereochemistry. LC-MS for C₂₂H₂₈F₆N₂O₄ [M+H]⁺ calculated: 499.20, found443.05 (loss of the t-butyl group). Higher eluting diastereoisomer: ¹HNMR (CDCl₃, 500 MHz): 7.76 (s, 1H), 7.75 (s, 2H), 4.86 (bs, 1H), 4.62(dd, J=15.6, 6.4 Hz, 1H), 4.49 (dd, J=15.6, 5.0 Hz, 1H), 3.98 (m, 1H),3.82 (dd, J=12.6, 6.2 Hz, 1H), 2.41 (m, 2H), 2.01 (m, 1H), 1.79 (dd,J=13.7, 6.6 Hz, 1H), 1.64 (m, 1H), 1.41 (s, 9H), 1.32 (m, 1H), 1.17 (d,J=6.18 Hz, 3H).Step P

A solution of the of the higher eluting diastereoisomer from theprevious step (92 mg, 0.19 mmol) in dichloromethane (3 mL) was treatedwith TFA (3 mL) and the resulting mixture was stirred at roomtemperature for 2 h. The volatiles were removed in vacuo to yield 86.9mg (93%) of the crude product. LC-MS for C₁₇H₂₀F₆N₂O₂ [M+H]⁺ calculated:399.14, found 399.15. A similar procedure was applied to the lowereluting BOC-protected amine described in the previous step to obtain therespective amine.Step G

The amide trifluoroacetate, the preparation of which was described inthe previous step (87 mg, 0.17 mmol), tetrahydro-4H-pyran-4-one (52 mg,0.52 mmol), crushed 4 Å molecular sieves (1.0 g) anddiisopropylethylamine were combined in dichloromethane and sodiumtriacetoxyborohydride (185 mg, 0.87 mmol) was added. The reactionmixture was stirred at room temperature overnight, after which time itwas quenched by pouring onto a saturated solution of sodium bicarbonate.The crude product was extracted with dichloromethane and the combinedorganic extracts were backwashed with brine, dried (anhydrous magnesiumsulfate) and the solvent was removed in vacuo. The residue (89.2 mg) wasfurther purified by preparative TLC (DCM:MeOH:ammonium hydroxide/90:9:1)to yield 65.8 mg of the pure product. LC-MS for C₁₂H₂₈F₆N₂O₃ [M+H]⁺calculated: 483.20, found 483.25. Similarly prepared was the finalproduct derived from the lower eluting BOC-protected amine as describedin Step E of this Example.

EXAMPLE 88

This compound was synthesized according to the procedure described inExample 63, except that 1-hydroxymethylcyclopentylamine was used insteadof Intermediate 2. The respective single diastereoisomers were separatedby preparative TLC. LC-MS for C₂₂H₃₂F₃N₃O₃ [M+H]⁺ calculated 444.24,found 444.24.

EXAMPLE 89

Step A

3-Cyano-2-fluorobenzotrifluoride (2.0 g, 11 mmol) was combined withhydrazine monohydrate (10 mL, 200 mmol) in n-butyl alcohol (40 mL) andheated to reflux. After 2.5 h the reaction was cooled to roomtemperature and concentrated to dryness to give 2.2 g of a white solid.ESI-MS calc. for C8H6F3N3: 201; found 202 (M+H).Step B

To a cooled (0° C.) solution of Intermediate 42 (63 mg, 0.18 mmol) inDCM (5 mL) was added oxalyl chloride (54 μL, 0.53 mmol) and 1 drop ofDMF and the resulting solution was allowed to warm to room temperature.After 2 h the reaction was concentrated to dryness and dried for 1.5 hunder high vacuum. The resulting acid chloride was dissolved in DCM (5mL) and added dropwise to a stirred solution of the product from Step A(55 mg, 0.25 mmol) in triethylamine (10 mL). After 30 minutes thereaction was concentrated under reduced pressure and the crudeintermediate was dissolved in ethyl alcohol and treated with sodiumborohydride (20 mg). After 20 h at room temperature the reaction wasconcentrated to dryness and the product was purified by reverse phaseHPLC (C18, 25-100% MeCN/H₂O) and converted to its hydrochloride salt bythe addition of hydrogen chloride (2 N solution in ethyl ether) to give1.3 mg of product. ESI-MS calc. for C22H29F3N4O2: 438; found 439 (M+H).

EXAMPLE 90

To a mixture of Intermediate 16 (0.95 g, 2.2 mmol) in dichloroethane (10mL) was added successively Intermediate 5 (0.276 g, 2.42 mmol), DIEA(0.31 g, 2.4 mmol) and 4 Å powdered molecular sieves (2.0 g) and theresulting mixture was stirred for 30 minutes at room temperature. To thestirred mixture was added sodium triacetoxyborohydride (0.7 g, 3 mmol)and the reaction was stirred at room temperature for 24 h. The mixturewas filtered and the filtrate was stirred with a saturated solution ofsodium bicarbonate (2.0 mL) for 10 minutes and then taken in ethylacetate (50 mL). The organic layer was washed with brine, dried(anhydrous magnesium sulfate), concentrated, and purified by flashcolumn chromatography (hexane:EtOAc/1:1+15% MeOH), to give the titlecompound (1.02 g, 94%) as a mixture of diastereoisomers. PreparativeHPLC (ChiralCel OD column) was used to facilitate further separation ofdiastereoisomers. Thus, 60 mg of the above mixture was separated(elution with hexane and 5% ethanol) to yield 4 mg, 8 mg and 18 mg ofthree isomers shown below. All three isomers were then transformed tothe HCl salts.

Following Example 90 and starting from other appropriate intermediates,a variety of compounds were prepared. Reverse phase preparative HPLCaccomplished the separation of selected compounds, which weresubsequently transformed to the HCl salts. Their structures (Examples 90to 131) and MS characteristics are listed in the Table 4. TABLE 4(Examples 90 to 131)

Molecular Calculated Found Ex. R Formula [M + H⁺] [M + H⁺] 90

C₂₄H₃₂F₆N₂O₂ 495.24 495.30 91

C₂₇H₃₁F₆N₂O₂ 495.24 495.30 92

C₂₄H₃₂F₆N₂O₂ 495.24 495.30 93

C₂₅H₃₄F₆N₂O₂ 509.26 509.40 94

C₂₅H₃₄F₆N₂O₂ 509.26 509.40 95

C₂₅H₃₄F₆N₂O₂ 509.26 509.40 96

C₂₃H₂₉F₇N₂O₂ 499.22 499.22 97

C₂₃H₂₉F₇N₂O₂ 499.22 499.22 98

C₂₃H₂₉F₇N₂O₂ 499.22 499.20 99

C₂₄H₂₉F₉N₂O₃ 565.21 565.30 100

C₂₄H₂₉F₉N₂O₃ 565.21 565.30 101

C₂₆H₃₆F₆N₂O₂ 523.28 523.30 102

C₂₅H₃₄F₆N₂O₃ 509.26 509.20 103

C₂₅H₃₄F₉N₂O₂ 509.26 509.20 104

C₂₄H₂₉F₉N₂O₃ 549.26 549.40 105

C₂₆H₃₄F₆N₂O₄ 553.25 553.40 106

C₂₆H₃₄F₆N₂O₄ 525.22 525.30 107

C₂₃H₃₀F₆N₂O₂ 481.23 481.20 108

C₂₃H₃₀F₆N₂OS 497.21 497.20 109

C₂₃H₃₀F₆N₂O₃S 529.20 529.20 110

C₂₄H₃₂F₆ ₂O₂ 495.24 495.30 112

C₂₂H₂₉F₆N₂O₂ 497.21 467.20 113

C₂₂H₂₉F₆N₂O₂ 467.21 467.20 114

C₂₂H₃₀F₆N₂OS 483.19 483.20 115

C₂₁H₂₆F₆N₂O₂ 453.20 453.15 116

C₂₄H₃₂F₆N₂O 479.25 479.30 117

C₂₅H₃₄F₆N₂O 493.27 493.30 118

C₂₅H₃₄F₆N₂O 493.27 493.30 119

C₂₃H₃₀F₆N₂O 453.20 453.15 120

C₂₂H₂₈F₆N₂O 451.22 451.30 121

C₂₃H₃₀F₆N₂O 465.23 465.30 122

C₂₇H₃₀F₆N₂O 513.23 513.30 123

C₂₇H₃₀F₆N₂O 513.23 513.40 124

C₂₇H₃₀F₆N₂O₂ 529.23 529.30 125

C₂₈H₃₂F₆N₂O 527.25 527.30 126

C₂₄H₃₄F₆N₂O 493.27 493.30 127

C₂₄H₃₂F₆N₂O₂ 495.24 495.40 128

C₂₅H₃₂F₆N₂O₂ 495.24 495.40 129

C₂₅H₃₂F₆N₂O₄ 539.23 539.30 130

C₂₄H₃₂F₆N₂O₃ 511.24 511.30 131

C₂₃H₃₂F₆N₂O 480.24 480.30

EXAMPLE 132

To a mixture of Intermediate 14 (0.5 g, 1.1 mmol) in dichloroethane (20mL) was added successively intermediate 25 (0.41 g, 3.2 mmol), DIEA(0.31 g, 2.42 mmol) and 4 Å powdered molecular sieves (2.0 g) and themixture was stirred for 30 minutes at room temperature. To this stirredmixture was added sodium triacetoxyborohydride (0.45 g, 2.1 mmol) andthe resultant mixture was stirred at room temperature for 24 h. Themixture was filtered and the filtrate was stirred with a saturatedsolution of sodium bicarbonate (2.0 mL) for 10 minutes and then taken inethyl acetate (50 mL). The solvent layer was washed with brine, dried(anhydrous magnesium sulfate), concentrated, and purified by flashcolumn chromatography. Eluting with DCM+10% MeOH, gave the titlecompound (0.42 g, 75%) as a mixtures of diastereomers. Reverse phaseHPLC (ChiralCel OD column) was used to facilitate further separation ofthe diastereomers discussed above. Elution with heptane containing 8%ethanol gave two major compounds 74 mg and 132 mg of two isomers shownbelow. Both isomers were then transformed to the HCl salt.

Following Example 132 and starting from other appropriate intermediates,a variety of compounds were prepared and are shown in Table 5.Preparative reverse phase HPLC accomplished the separation of selectedcompounds, which were subsequently transformed to the HCl salts. Theirstructures (Examples 132 to 140) and MS characteristics are summarizedin Table 5. TABLE 5 (Examples 90 to 131)

Molecular Calculated Found Ex. R Formula [M + H⁺] [M + H⁺] 132

C₂₅H₃₄F₆N₂O₃ 525.26 525.40 133

C₂₅H₃₄F₆N₂O₃ 525.26 525.40 134

C₂₃H₂₉F₇N₂O₃ 515.21 515.40 135

C₂₃H₂₉F₇N₂O₃ 515.21 515.40 136

C₂₆H₃₄F₆N₂O₃ 537.26 537.40 137

C₂₃H₃₀F₆N₂O₃ 497.22 497.20 138

C₂₃H₃₀F₆N₂O₂S 513.20 513.20 139

C₂₄H₃₂F₆N₂O₄ 527.23 — 140

C₂₄H₃₂F₆N₂O₂S 527.22 527.40

EXAMPLE 141

To a solution of amine from Example 91 (0.028 g, 0.057 mmol) in MeOH(2.0 mL) was added formalin (40 □L, 37% solution in water) followed bysodium cyanoborohydride (0.010 g, 0.17 mmol) and the reaction wasstirred at room temperature for 16 h. After the evaporation of thevolatiles, the crude was purified by silica column chromatography.Elution with hexane:EtOAc (1;1)+ 2% MeOH gave the desired product.

Starting from Examples 92, 96 and 97 and following the procedure givenfor Example 141 the products listed below were obtained. The synthesizedcompounds (Examples 141 to 144) and MS characteristics are summarized inTable 6. TABLE 4 (Examples 90 to 131)

Molecular Calculated Found Ex. R Formula [M + H⁺] [M + H⁺] 141

C₂₅H₃₄F₆N₂O₂ 509.26 509.30 142

C₂₅H₃₄F₆N₂O₂ 509.26 509.30 143

C₂₄H₃₁F₇N₂O₂ 513.24 513.30 144

C₂₄H₃₁F₇N₂O₂ 513.24 513.30

EXAMPLE 145

Step A

To intermediate 10 (0.27 g, 1.0 mmol) and 3-fluoro-5-trifluromethylbenzylamine (0.23 g, 1.2 mmol) in DCM (8.0 mL) was added PyBOP (0.75 g,1.5 mmol) and the resulting mixture was stirred at room temperature for24 h. The reaction mixture was quenched with water and the solvent layerwas washed with brine, dried (anhydrous magnesium sulfate),concentrated, and purified by flash column chromatography. Eluting withhexane:EtOAc (10:1) gave the title compound (0.14 g, 32%).Step B:

To a solution of the product from Step A (0.14 g) in EtOAc (4.0 mL) wasadded a saturated solution of HCl in EtOAc (1.0 mL) and the mixturestirred. After 1 h at room temperature, the volatiles were removed undervacuum to leave behind the title compound (0.12 g, 100%) as a whitesolid. LC-MS for C₁₇H₂₂F₄N₂O [M+H]⁺ calculated 346.17, found 346.2.Step C

A mixture of the intermediate from Step B (0.035 g, 0.090 mmol),intermediate (0.018 g, 0.14 mmol), DIEA (0.013 g, 0.101 mmol) and 4 Åpowdered molecular sieves (0.1 g) in dichloroethane (2 mL) was stirredfor 30 minutes at room temperature. To the stirred mixture was addedsodium triacetoxyborohydride (0.029 g, 0.14 mmol) and the resultantmixture was stirred at room temperature for 24 h. The mixture wasfiltered and the filtrate was evaporated and purified by flash columnchromatography. Eluting with hexane:EtOAc (1:1)+ 10% MeOH, gave Example145 as a mixture of diastereomers which was subsequently transformed toits HCl salt (0.018 g). LC-MS for C₂₄H₃F₄N₂O₂ [M+H]⁺ calculated 459.26,found 459.4.

EXAMPLE 146

Step A

A mixture of 3-bromo-5-trifluoromethylbenzonitrile (1.0 g, 4.2 mmol),triethyl orthoformate (1.3 mL, 8.1 mmol) and sodium azide (490 mg, 7.5mmol) in glacial acetic acid (10 mL) was heated to reflux for 8 h. Thereaction was poured onto crushed ice and extracted 4 times with ethylether. The combined organic layers where washed with brine, dried overMgSO₄, filtered and concentrated under reduced pressure. The residue waspurified by medium pressure liquid chromatography (silica gel, 0-60%EA/hexanes) to give 510 mg of product (41%). ESI-MS calc. forC8H4BrF3N4: 292; found 293 (M+H).Step B

The product from Step A (440 mg, 1.3 mmol) was combined withtetrakis(triphenylphosphine) palladium (90 mg, 0.078 mmol) and zinccyanide (300 mg, 2.6 mmol) in DMF (deoxygenated, 3 mL) and heated toreflux. After 22 h, the reaction was cooled to room temperature andpartitioned between ethyl ether and aqueous 2 N NH₄OH. The organic layerwas washed twice with 2 N NH₄OH, then with brine, dried over MgSO₄,filtered and concentrated under reduced pressure to give 340 mg of aproduct which was used directly in the next step. ESI-MS calc. forC9H4F3N5: 239; found 240 (M+H).Step C

The product from Step B (340 mg) was dissolved in THF (10 mL) andtreated with borane (1.0 M solution in THF, 7.0 mL, 7.0 mmol). After 20h at room temperature, the reaction was quenched with a 1% hydrogenchloride solution in methanol (20 mL) and heated to 50° C. After 3 h.the reaction was concentrated under reduced pressure and the residue wasre-dissolved in a 1% hydrogen chloride solution in methanol (20 mL).After 18 h the solution was concentrated under reduced pressure and theresulting residue was dissolved in aqueous 1N HCl, and washedsuccessively with DCM and ethyl ether. The aqueous layer wasconcentrated under reduced pressure to give 300 mg of a whitehydrochloride salt (73% over 2 steps). ESI-MS calc. for C9H8F3N5: 243;found 244 (M+H).Step D

Intermediate 17 (700 mg, 2.6 mmol) was dissolved in DCM (10 mL) andtreated with oxalyl chloride (500 μL, 5.5 mmol) and 1 drop of DMF. After2 h at room temperature the reaction was concentrated to dryness anddried for 1.5 under high vacuum. 35 mg of this acid chloride (0.12 mmol)was dissolved in DCM (1 mL) and added dropwise to a stirred solution ofthe product from Step C (16 mg, 0.057 mmol) in triethylamine (1 mL).After 3.5 h the reaction was concentrated under reduced pressure andpassed through a Spe-ed SCX column, washing with methanol and elutingwith 2 M NH₃ solution in methanol. This crude product was furtherpurified by reverse phase HPLC (C18, 25-100% MeCN/H₂O) and converted toits hydrochloride salt by addition of hydrogen chloride (2 N solution inethyl ether) to give 2.5 mg of a white solid (8%). ESI-MS calc. forC24H33F3N6O2: 494; found 495 (M+H).

EXAMPLE 147

Step A

To a cooled (0° C.) solution of 2-fluoro-5-trifluoromethylbenzonitrile(5.23 g, 27.7 mmol) in 140 mL of THF was added dropwise at a rapid pacea suspension of potassium t-butoxide (3.88 g, 34.6 mmol) in 35 mL ofTHF. The reaction mixture was permitted to slowly warm to roomtemperature and stir overnight. The reaction mixture was concentratedunder reduced pressure then ethyl ether and 1 N HCl solution were addedand the layers were separated. The ethereal layer was washed withsaturated NaHCO₃ solution, then brine, dried over anhydrous MgSO₄,filtered, and concentrated. Purification by MPLC (silica, 25% ethylacetate/hexane) afforded 5.25 g (78%) of a white crystalline solid. ¹HNMR (CDCl₃, 500 MHz): 7.84 (d, J=2.0 Hz, 1H), 7.73 (dd, J=8.5, 2.0 Hz,1H), 7.27 (d, J=9.0 Hz), 1.55 (s, 9H).Step B

To a solution of the nitrile prepared as described in Step A (7.6 g, 31mmol) in ethanol (100 mL) was added ammonium hydroxide solution (28-30%,25 mL) and Raney® 2800 nickel (slurry in water, ˜3.5 g). The resultingmixture was agitated under 50 psi of hydrogen gas for 24 h using a Parrapparatus. The reaction mixture was then filtered through celite washingwith ethanol and then water. The filtrate was concentrated to drynessunder reduced pressure and the residue so obtained was purified by flashchromatography [silica, 5 to 10% gradient (1% increments) of (10%ammonium hydroxide solution (28-30%)/methanol) in DCM] to afford 5.5 g(71%) of desired amine as a colorless oil which crystallized uponstorage in the freezer. ¹H NMR (CDCl₃, 500 MHz): 7.56 (d, J=2.0 Hz, 1H),7.44 (dd, J=8.5, 2.0 Hz, 1H), 7.12 (d, 8.5 Hz, 1H), 3.90 (s, 2H), 2.70(bs, 2H), 1.51 (s, 9H).Step C

Intermediate 42 (10 g, 39 mmol) was combined withtetrahydro-4H-pyran-4-one (4.3 g, 43 mmol), triethylamine (5.4 mL, 39mmol), 4 Å powdered molecular sieves (5 g), and sodiumtriacetoxyborohydride (41 g, 190 mmol) in 200 mL DCM. The reactionmixture was stirred at room temperature for 4 days, then filteredthrough celite, diluted with DCM, and washed with saturated NaHCO₃solution twice and then once with brine. The organic layer was driedover anhydrous MgSO₄, filtered and concentrated to give 10 g of product(85%). ESI-MS calc. for C18H25NO3: 303; found 304 (M+H).Step D

The product from Step C (10 g, 33 mmol) was combined with formaldehyde(37% solution in water) (27 mL, 330 mmol), 4 Å powdered molecular sieves(10 g), and sodium triacetoxyborohydride (35 g, 170 mmol) in 350 mL DCM.The reaction mixture was stirred at room temperature for 4 h, thenfiltered through celite, diluted with DCM, and washed with saturatedNaHCO₃ and brine. The organic layer was dried over anhydrous MgSO₄,filtered and concentrated to give 8.4 g of product (81%). ESI-MS calc.for C19H27NO3: 317; found 318 (M+H).Step E

The product from Step D (1.1 g, 3.4 mmol) was hydrogenolyzed overPd(OH)₂ (20%, 110 mg) in methanol under a hydrogen filled balloon. After90 minutes the reaction was filtered through celite and concentratedunder reduced pressure to give 900 mg of product.Step F

To a stirred solution of the product from Step E (50 mg, 0.22 mmol) and1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (62 mg, 0.32 mmol) in DCM(10 mL), was added the product from Step B (80 mg, 0.32 mmol). Thereaction was stirred at room temperature for 18 h before being dilutedwith DCM and washed with saturated aqueous sodium bicarbonate and brine.The organic layer was dried over MgSO₄, filtered and concentrated underreduced pressure. The product was purified preparative TLC (silica gel,0.3% NH₄OH (3.7% MeOH (97% DCM) to give 68 mg of a colorless oil (67%).1.3 mg of this oil was converted to its hydrochloride salt by additionof hydrogen chloride (2 N solution in ethyl ether) to give 1.4 mg of awhite solid. ESI-MS calc. for C24H35F3N2O3: 456; found 457 (M+H).

EXAMPLE 148

The free base from Example 147 (66 mg, 0.14 mmol) was dissolved intrifluoroacetic acid (3 mL) and treated with 3 drops of water. After 72h at room temperature, the reaction was concentrated under reducedpressure and converted to its hydrochloride salt by the addition ofhydrogen chloride (2 N solution in ethyl ether) to give 62 mg of a whitesolid. ESI-MS calc. for C20H27F3N2O3: 400; found 401 (M+H).

EXAMPLE 149

Step A

3-Cyano-4-fluorobenzotrifluoride (2.0 g, 11 mmol) was combined withhydrazine monohydrate (10 mL, 200 mmol) in n-butyl alcohol (40 mL) andheated to reflux. After 2 h the reaction was cooled to room temperatureand concentrated to dryness to give 2.6 g of a white solid. ESI-MS calc.for C8H6F3N3: 201; found 202 (M+H). ¹H NMR (CDCl₃, 500 MHz): δ 8.18 (s,1H), 7.45 (d, 1H), 7.35 (d, 1H), 5.60 (m, 3H).Step B

Intermediate 42 (200 mg, 0.6 mmol) was dissolved in DCM (5 mL) and THF(10 mL) and was treated with oxalyl chloride (110 μL, 1.2 mmol) and 1drop of DMF. After 2 h at room temperature the reaction was concentratedto dryness and dried for 1.5 h under high vacuum. 100 mg of this acidchloride (0.3 mmol) was dissolved in DCM (5 mL) and added dropwise to astirred solution of the product from Step A (180 mg, 0.90 mmol) intriethylamine (3 mL). After 72 h the reaction was concentrated underreduced pressure and the crude product was dissolved in a solution ofTHF (3 mL) and methanol (3 mL) and treated with a solution of lithiumhydroxide (20 mg) in water (3 mL). This solution was stirred at roomtemperature for 72 h and concentrated to dryness. This crude product wasfurther purified by reverse phase HPLC (C18, 25-100% MeCN/H₂O) andconverted to its hydrochloride salt by the addition of hydrogen chloride(2 N solution in ethyl ether) to give 0.8 mg of product. ESI-MS calc.for C22H29F3N4O2: 438; found 439 (M+H).

EXAMPLE 150

The dicarbonyl Intermediate 39 (36 mg, 0.23 mmol) was combined withoptically pure amine Intermediate 16 (100 mg, 0.231 mmol), triethylamine(32 μL, 0.23 mmol), 4 Å powdered molecular sieves (˜100 mg), and sodiumtriacetoxyborohydride (245 mg, 1.16 mmol) in DCM (3 mL). The resultingmixture was stirred at room temperature for 16 h. The reaction mixturewas filtered through a celite plug, washing with more DCM. The filtratewas washed with saturated NaHCO₃ solution, then brine. The organic layerwas dried over anhydrous MgSO₄, filtered, and concentrated. Purificationby flash chromatography (silica, gradient from 0.25/2.25/97.5 to 1/9/90of NH₄OH/methanol/DCM) gave three bands, each having the correct massfor the product (4 possible isomers separated into three bands). Mixedfractions obtained from the first purification were subjected to furtherpurification by preparative TLC (silica, 0.6/5.5/94 ofNH₄OH/methanol/DCM) and the bands were combined with the correspondingbands purified by flash chromatography giving 34 mg, 42 mg, and 12 mg ofthe 1^(st), 2^(nd), and 3^(rd) bands to elute, respectively. Analysis bychiral HPLC (ChiralPak AD and OD columns) indicated that the first twobands were single isomers, while the third band was a mixture of twoisomers. All three products were converted to their HCl salts bydissolving in DCM (˜1 mL), adding excess 4 N HCl in dioxane (˜6 drops)and concentrating to give white solids.

Spot 1, single isomer: ESI-MS calc. for C26H34F6N2O2: 520; Found: 521(M+H).

Spot 2, single isomer: ESI-MS calc. for C26H34F6N2O2: 520; Found: 521(M+H).

Spot 3, two isomers: ESI-MS calc. for C26H34F6N2O2: 520; Found: 521(M+H).

EXAMPLE 151

The dicarbonyl Intermediate 39 (34 mg, 0.22 mmol) was combined withoptically pure amine Intermediate 14 (100 mg, 0.223 mmol), triethylamine(31 μL, 0.22 mmol), 4 Å powdered molecular sieves (˜200 mg), and sodiumtriacetoxyborohydride (236 mg, 1.12 mmol) in DCM (4 mL). The resultingmixture was stirred at room temperature for 24 h. The reaction mixturewas filtered through a celite plug, washing with methanol. The filtratewas concentrated. Saturated NaHCO₃ solution (30 mL) and methanol (˜5 mL)was added and the reaction mixture was stirred at 50° C. for 0.5 h (tobreak up boron complex with product as seen in HPLC-MS). The mixture waspartially concentrated to remove methanol, and then was extracted twicewith DCM. The combined organic layers were washed with brine, dried overMgSO₄, filtered, and concentrated. Purification by preparative TLC(silica, 0.7/6.3/93 of NH₄OH/methanol/DCM) permitted separation of fourbands, each with a mass corresponding to product (resolved all fourpossible isomers). The top spot (spot 1) required further purificationby reverse phase HPLC (YMC column) to eliminate a side product andobtain the pure isomer. All four pure isomers were converted to theirHCl salts by dissolving in DCM (˜1 mL), adding excess 4 N HCl in dioxane(˜6 drops) and concentrating to give white solids.

Spot 1: ESI-MS calc. for C26H34F6N2O3: 536; Found: 537 (M+H).

Spot 2: ESI-MS calc. for C26H34F6N2O3: 536; Found: 537 (M+H).

Spot 3: ESI-MS calc. for C26H34F6N2O3: 536; Found: 537 (M+H).

Spot 4: ESI-MS calc. for C26H34F6N2O3: 536; Found: 537 (M+H).

EXAMPLE 152

The dicarbonyl Intermediate 40 (33 mg, 0.23 mmol) was combined withoptically pure amine Intermediate 16 (100 mg, 0.231 mmol), triethylamine(32 μL, 0.23 mmol), 4 Å powdered molecular sieves (˜200 mg), and sodiumtriacetoxyborohydride (244 mg, 1.16 mmol) in DCM (20 mL). The resultingmixture was stirred under nitrogen at room temperature for 16 h. Thereaction mixture was filtered through a celite plug, washing with moreDCM. The filtrate was washed with saturated NaHCO₃ solution, then brine.The organic layer was dried over anhydrous MgSO₄, filtered, andconcentrated. Purification by flash chromatography (silica, gradientfrom 0.1/0.9/99 to 1.5/13.5/85 of NH₄OH/methanol/DCM) gave a single spotpresumed to be the 2 cis-product isomers. Purification by chiral HPLC(ChiralPak OD column, 5% ethanol/hexane) afforded two pure singleisomers: peak 1 gave 18 mg, and peak 2 gave 16 mg. ESI-MS calc. forC25H32F6N2O2: 506; Found: 507 (M+H).

EXAMPLE 153

The dicarbonyl Intermediate 40 (210 mg, 1.48 mmol) was combined withoptically pure amine Intermediate 14 (610 mg, 1.48 mmol), triethylamine(205 μL, 1.48 mmol), 4 Å powdered molecular sieves (˜1 g), and sodiumtriacetoxyborohydride (1.57 g, 7.40 mmol) in DCM (50 mL). The resultingmixture was stirred at room temperature for 3 days. The reaction mixturewas filtered through a celite plug, washing with methanol. The filtratewas concentrated. Saturated NaHCO₃ solution (100 mL) was added and thereaction mixture was agitated at 50° C. for 3 h (to break up boroncomplex with product as seen in HPLC-MS). The aqueous mixture wasextracted six times with DCM, then the organic layers were combined andwashed with brine, dried over MgSO₄, filtered, and concentrated to give653 mg of crude product. Separation of the two cis-isomers and furtherpurification was accomplished by a combination of preparative TLC(silica, 0.5/4.5/95 of NH₄OH/methanol/DCM) and chiral HPLC (ChiralPak ODcolumn, 7% ethanol/hexane) afforded the resolved single cis-isomers,which were converted to their HCl salts by dissolving in DCM, addingexcess 4 N HCl in dioxane and concentrating to give white solids (180.3mg of peak 1 off of the OD column, and 147.6 mg of peak 2). ESI-MS calc.for C25H32F6N2O3: 522; Found: 523 (M+H).

EXAMPLE 154

The ketone Intermediate 41 (43 mg, 0.31 mmol) was combined withoptically pure amine Intermediate 16 (133 mg, 0.307 mmol), triethylamine(43 μL, 0.31 mmol), 4 Å powdered molecular sieves (˜200 mg), and sodiumtriacetoxyborohydride (260 mg, 1.23 mmol) in DCM (4 mL). The resultingmixture was stirred at room temperature overnight. The reaction mixturewas filtered through a celite plug, washing with more DCM. The filtratewas washed with saturated NaHCO₃ solution, then brine. The organic layerwas dried over anhydrous MgSO₄, filtered, and concentrated. Purificationby preparative TLC (silica, 5% of 1:9 NH₄OH/methanol in DCM) gave twobands corresponding to product (74 mg top spot, 60 mg bottom spot).

Top spot-ESI-MS calc. for C26H34F6N2O2: 520; Found: 521 (M+H).

Bottom spot-ESI-MS calc. for C26H34F6N2O2: 520; Found: 521 (M+H).

EXAMPLE 155

Step A

A solution of ethyl cyanoacetate (40.9 g, 0.361 mol) in 400 mL DMF wascooled to 0° C. and treated under a steady stream of N₂ with lithiumhydride (7.18 g, 0.903 mol) in multiple portions. After hydrogenevolution subsided, cis-1,4-dichloro-2-butene (51.9 g, 0.415 mol) wasadded dropwise by addition funnel. The reaction became very thick duringthe addition, requiring the addition of 200 mL of DMF to aid instirring. The reaction mixture was permitted to warm to room temperatureand was stirred for 1 h. The reaction mixture was then poured into a 1:1mixture of water/ice, which was in turn extracted twice with ether. Theethereal layers were combined and washed five times with water, and oncewith brine. The ethereal phase was then dried over MgSO₄, filtered andconcentrated. The resulting crude product was distilled using a shortpath distillation apparatus (1 mm Hg, bath temperature=100° C., headtemperature=75° C.), giving 25.8 g of the desired product (43%). ¹H NMR(CDCl₃, 500 MHz): 5.70 (s, 2H), 4.27 (q, J=7 Hz, 2H), 3.10 (m, 4H), 1.34(t, J=7 Hz, 3H).Step B

A solution of the cyclopentene prepared in Step A above (17.5 g, 0.106mol) in 100 mL of THF was cooled to −78° C. and treated with BH₃-THF(1.0 M solution in THF, 63.5 mL, 64 mmol) dropwise. The reaction mixturewas stirred at −78° C. for 0.5 h, then warmed to room temperature andstirred for an additional 1 h. TLC indicated that the reaction wasincomplete so the mixture was cooled back to −78° C. and treated withmore BH₃-THF solution (1.0 M solution in THF, 42 mL, 42 mmol). Thereaction mixture was then warmed to room temperature and stirred for 2h. After storing overnight in a freezer, the reaction mixture wasconcentrated at room temperature and redissolved in DCM (500 mL). Thenwhile stirring with an overhead mechanical stirring apparatus, premixedPCC (137 g, 0.635 mol) and magnesium sulfate (130 g) were added inportions over 15 minutes. The resulting exotherm was controlled with anice bath. After stirring at room temperature for 3 h, the reactionmixture was filtered through a 3″ plug of silica, washing the remainingsolids three times with acetone. The filtrate was concentrated andfiltered a second time through a 3″ silica plug washing through with 50%ethyl acetate/hexane. The filtrate was concentrated and the residue waspurified by flash chromatography (silica, 50% ethyl acetate/hexane)giving 4.63 g (24%) of product. ¹H NMR (CDCl₃, 500 MHz) δ 4.35 (q, J=8.5Hz, 2H), 2.94 (d, J=23 Hz, 1H), 2.78 (d, J=23 Hz, 1H), 2.51-2.70 (m,4H), 1.38 (t, J=9 Hz, 3H).Step C

The ketone intermediate from Step B above (1.02 g, 5.64 mmol) wascombined with Intermediate 2 (931 mg, 6.77 mmol), sodiumtriacetoxyborohydride (4.78 g, 22.6 mmol), triethylamine (0.94 mL, 6.8mmol), and 4 Å powdered molecular sieves (˜2 g) in 30 mL of DCM. Thereaction mixture was permitted to stir at room temperature for 4 days.Then 37% aqueous formaldehyde (4.58 g, 56.4 mmol) was added, followed bymore 4 Å powdered molecular sieves (˜5 g). After stirring the mixturefor five minutes, more sodium triacetoxyborohydride (5 g, 23 mmol) wasadded and the resulting mixture was stirred for 2.5 h. The reactionmixture was then filtered through celite, washing with DCM. The filtratewas then washed with saturated NaHCO₃ solution, water, and brine, driedover anhydrous MgSO₄, filtered, and concentrated. Purification by flashchromatography (silica, 10% methanol/DCM) afforded 1.12 g of targetamine (75% yield). ESI-MS calc. for C15H24N2O3: 280; Found: 281 (M+H).Step D

A solution of the aminoester from Step C above (1.07 g, 3.82 mmol) in1:1 THF/methanol (16 mL) was treated with a solution of LiOH.H₂O (0.80g, 19 mmol) in water (8 mL). The resulting mixture was stirred at roomtemperature for 2 h, then was neutralized with 2 N HCl solution, andpartially concentrated to remove the organic solvents. Attempts toextract the product into chloroform failed as the product remained inthe aqueous layer. Therefore the aqueous mixture was concentrated todryness (2.06 g crude mixture) and used as is in the following step.ESI-MS calc. for C13H20N2O3: 252; Found: 253 (M+H).Step E

A solution of the crude amino-acid from Step D above (˜3.17 mmol) and3,5-Bis(trifluoromethyl)-benzylamine hydrochloride (1.33 g, 4.76 mmol)in DCM was treated with EDC (1.22 g, 6.34 mmol). The resulting reactionmixture was stirred at room temperature over night. HPLC-MS analysisindicated low conversion. 4 N HCl in 1,4-dioxane (0.79 mL, 3.2 mmol) wasadded followed by DMF (10 mL) and the reaction mixture was stirred foranother 48 h. The reaction mixture was then partially concentrated toremove the DCM and partitioned between EtOAc and saturated NaHCO₃solution. The aqueous layer was extracted again with ethyl acetate andthe combined organic layers were washed with water five times and oncewith brine. The organic layer was then dried over anhydrous MgSO₄,filtered, and concentrated. Purification by preparative TLC allowedseparation of the cis (top spot) and trans (bottom spot) isomers,although approximately ⅓ of the material was lost accidentally. 198 mgOf the desired cis isomer was obtained as a mixture of two enantiomers.ESI-MS calc. for C22H25F6N3O2: 477; Found: 478 (M+H).

EXAMPLE 156

To a solution of the final product in Example 155 (87.6 mg, 0.183 mmol)in DMSO (1 mL) was added K₂CO₃ (5 mg) followed by 30% H₂O₂ solution (24μL). The resulting mixture was stirred at room temperature for 30minutes, then was quenched with excess 10% Na₂SO₃ solution. The mixturewas then extracted three times with ethyl acetate and the combinedorganic layers were washed successively with 10% Na₂SO₃ solution, water(four times) and brine. The organic layer was then dried over anhydrousMgSO₄, filtered, and concentrated to afford 76.6 mg of crude product.Purification by preparative TLC (silica, 8% of 1/9 NH₄OH/methanol inDCM) gave 49.1 mg of target compound.

EXAMPLE 157

A solution of the final product from Example 156 (47.6 mg, 0.0961 mmol)in N,N-dimethylformamide dimethylacetal (1.5 mL) was stirred at 120° C.for 3 h, then was concentrated and stored under vacuum until the nextday. The residue was dissolved in AcOH (1 mL), hydrazine hydrate (6 mg)was added, and the mixture was stirred at 90° C. for 3 h. The reactionmixture was concentrated and the crude product was purified by reversephase HPLC, followed by preparative TLC (silica, 10% of 1/9NH₄OH/methanol in DCM) to give the target compound as a mixture of twoenantiomers. ESI-MS calc. for C23H27F6N5O2: 519; Found: 520 (M+H).

EXAMPLE 158

Step A

The final product from Example 155 (87 mg, 0.18 mmol) was combined withtriethylamine hydrochloride (75 mg, 0.55 mmol) and sodium azide (71 mg,1.1 mmol) in 1-methyl-2-pyrrolidinone (3 mL) and stirred at reflux for5.5 h, then at room temperature overnight. An aqueous workup failedbecause of the product's high aqueous solubility. The aqueous productmixture was therefore concentrated (1-2 mm Hg and ˜70° C. were requiredto remove the 1-methyl-2-pyrrolidinone). The crude product was purifiedby preparative TLC (silica, 30% methanol/DCM) to give 58 mg of thetetrazole product. ESI-MS calc. for C22H26F6N6O2: 520; Found: 521 (M+H).Step B

The tetrazole intermediate prepared in Step A above (15.1 mg, 0.0290mmol) was combined with triphenylphosphine (19 mg, 0.073 mmol) andmethanol (3 μL, 0.07 mmol) in DCM under a nitrogen atmosphere. Diethylazodicarboxylate (12 μL, 0.073 mmol) was added and the reaction mixturewas stirred at room temperature overnight. The reaction mixture wasapplied onto an ion exchange column (Super Spe-ed benzenesulfonic SCX, 5g/35 mL from Applied Separations) and the column was rinsed with 10%methanol/ethyl acetate (50 mL) to remove neutral impurities. Then thecolumn was rinsed with 1:1 of 2 N NH₃ in methanol/DCM (40 mL) and thefiltrate was concentrated and further purified by preparative TLC(silica, 10% methanol in DCM) to give 8.9 mg of desired product (mixtureof two enantiomers). ESI-MS calc. for C23H28F6N6O2: 534; Found: 535(M+H).

EXAMPLE 159

Step A

Intermediate 8 (115 mg, 0.290 mmol) was combined with3-amino-1-N-t-butoxycarbonylpiperidine (87 mg, 0.44 mmol) and sodiumtriacetoxyborohydride (246 mg, 1.16 mmol) in 5 mL DCM and stirred atroom temperature overnight. The reaction mixture was diluted with DCMand washed with saturated NaHCO₃ solution, and brine, dried overanhydrous MgSO₄, filtered, and concentrated. Purification by preparativeTLC (silica, 8% of 1/9 NH₄—OH/methanol in DCM) allowed separation of twobands with the top band corresponding to a mixture of 4 cis-cyclopentylisomers (74 mg). ESI-MS calc. for C28H39F6N3O3: 579; Found: 580 (M+H).Step B

The product from Step A above (72 mg, 0.12 mmol) was dissolved in 5 mLof 4 N HCl in 1,4-dioxane and stirred at room temperature for 1.5 h. Thereaction mixture was then concentrated to afford 69.5 mg of product (asa mixture of four isomeric salts). ESI-MS calc. for C23H31F6N3O: 479;Found: 480 (M+H).Step C

A solution of the intermediate from Step B above (68 mg, 0.12 mmol) andtriethylamine (34 μL, 0.25 mmol) in 3 mL DCM was treated withmethanesulfonyl chloride (10 μL, 0.12 mmol) and stirred at roomtemperature for 1 h. The reaction mixture was concentrated and theresulting residue was purified by preparative TLC (silica, 8% of 1/9NH₄OH/methanol in DCM). The product was converted to its hydrochloridesalt by dissolving in DCM, adding excess 1 N HCl in ether (˜0.5 mL), andconcentrating to afford 55 mg of product salt. ESI-MS calc. forC24H33F6N3O3S: 557; Found: 558 (M+H).

EXAMPLE 160

Step A

To a solution of 4-chromanone (5.00 g, 33.7 mmol) in methanol (50 mL)was added concentrated HCl solution (28 mL, 340 mmol) and PtO₂ (0.5 g).The resulting mixture was agitated while under 50 psi of hydrogen gasfor 5 h using a Parr apparatus. The major products were those arisingfrom over reduction (for example, 3-cyclohexyl-1-propanol), howeverpurification by MPLC (silica, 50% ethyl acetate/hexane) did provide 136mg of desired product as a mixture of isomers.Step B

A solution of DMSO (0.244 mL, 3.43 mmol) in 2 mL DCM was added dropwiseto a cooled (−78° C.) solution of oxalyl chloride (0.150 mL, 1.72 mmol)in 20 mL of DCM. After stirring for an additional 3 minutes, the alcoholprepared as described in Step A above (134 mg, 0.858) was added dropwisein 2 mL of DCM. After 15 minutes, neat triethylamine (0.956 mL, 6.86mmol) was added dropwise. After 5 more minutes the reaction mixture waspermitted to warm to room temperature and stir for 1 h. The reactionmixture was then diluted with DCM and washed successively with 1 N HClsolution, saturated NaHCO₃ solution, and brine, dried over anhydrousMgSO₄, filtered, and concentrated. The resulting crude product waspurified by MPLC (silica, 50% ethyl acetate/hexane) to give 88 mg of theproduct as a 2:1 mixture of trans to cis isomers (see JOC (1974), 39,2040.). ¹H NMR (CDCl₃, 500 MHz): 4.30 (m), 3.81 (m, 1H from CHOH cisisomer), 3.75 (m), 3.23 (dt, J=10.5, 4.0 Hz, 1H from CHOH trans isomer),2.74 (m), 2.39 (m, 1H from cis isomer), 2.34 (m, 1H from trans isomer),2.22 (m), 2.10 (m), 2.03-1.95 (m), 1.88 (dq, J=12.5, 4.0 Hz), 1.84-1.75(m), 1.70-1.61 (m), 1.59-1.44 (m), 1.35-1.19 (m).Step C

The ketone from Step B (29.9 mg, 0.194 mmol) was combined withIntermediate 16 (76.3 mg, 0.176 mmol), triethylamine (25 μL, 0.18 mmol),and sodium triacetoxyborohydride (187 mg, 0.880 mmol) in 3 mL DCM andthe resulting mixture was stirred at room temperature for overnight. Thereaction mixture was diluted with DCM and washed with saturated NaHCO₃solution, then brine, dried over anhydrous MgSO₄, filtered, andconcentrated. The crude product was purified and separated into threeproduct bands (attributed to different isomer mixtures) by preparativeTLC (silica, 5% of 1/9 NH₄OH/methanol in DCM). The following amountswere collected: Top band 25 mg, middle band 18 mg, bottom band 6 mg. Allthree bands showed the correct mass as indicated below. All three bandsESI-MS calc. for C27H36F6N2O2: 534; Found: 535 (M+H).

EXAMPLE 161

Step A

To a mixture of 2,5-dihydrofuran (5.31 mL, 70.1 mmol) and Rh(OAc)₂ (1.55g, 7.01 mmol) in 250 mL of DCM was added via syringe pump under anitrogen atmosphere a solution of ethyl diazoacetate (8.85 g, 84.1 mmol)in 40 mL of DCM at a rate of 2 mmol/h. After the addition was completethe reaction mixture was filtered through celite and concentrated.Purification by MPLC (silica, 25% ethyl acetate/hexane) afforded 6.35 gof product which appeared by HNMR to have purely the trans cyclopropanestereochemistry. ¹H NMR (CDCl₃, 400 MHz) δ 4.14 (q, J=7.2 Hz, 2H), 3.94(d, J=8.8 Hz, 2H), 3.76 (d, J=8.4 Hz, 2H), 2.17 (m, 2H), 1.62 (t, J=3.2Hz, 1H), 1.28 (t, J=7.2 Hz, 3H).Step B

A solution of the ester prepared as described in Step A (5.35 g, 34.3mmol) in 80 mL of methanol was treated with a solution of LiOH.H₂O in 30mL of water. The reaction mixture was stirred overnight at roomtemperature. The reaction mixture was partially concentrated to removethe methanol and ˜32 mL of 4 N HCl in 1,4-dioxane was added whilecooling in an ice bath, bringing the pH to 5. The mixture was thenconcentrated almost to dryness and extracted twice with ethyl acetate.The combined organic layers were dried over anhydrous Na₂SO₄, filtered,and concentrated to give 2.41 g of product which required no furtherpurification. ¹H NMR (CDCl₃, 400 MHz): 3.97 (d, J=8.8 Hz, 2H), 3.78 (d,J=8.8 Hz, 2H), 2.25 (m, 2H), 1.64 (t, J=3.2 Hz, 1H).Step C

The acid from Step B (1.40 g, 10.9 mmol) was combined withdiphenylphosphoryl azide (2.59 mL, 12.0 mmol) and triethylamine (3.19mL, 22.9 mmol) in 90 mL of t-butanol and stirred at reflux for 3 days(timer failed to turn off). The reaction mixture was concentrated andthe residue was dissolved in ethyl acetate and washed with 1 N HClsolution, saturated NaHCO₃ solution, and brine, dried over MgSO₄,filtered, and concentrated. Purification by MPLC (silica, 65% ethylacetate/hexane) gave 1.14 g of the desired product. 1H NMR (CDCl₃, 400MHz): 4.65 (br s, 1H), 3.98 (d, J=8.0 Hz, 2H), 3.72 (d, J=8.4 Hz, 2H),2.41 (s, 1H), 1.78 (s, 2H), 1.46 (s, 9H).Step D

A solution of the product from Step C (790 mg, 3.96 mmol) in 4 N HCl in1,4-dioxane was stirred at room temperature for 0.5 h then concentratedto give 537 mg of amine hydrochloride product. 1H NMR (CD₃OD, 500 MHz):3.94 (d, J=9.0 Hz, 2H), 3.69 (d, J=9.0 Hz, 2H), 2.38 (br t, J=2.5 Hz,1H), 2.07 (m, 2H).Step E

A solution of the amine salt prepared in Step D (62 mg, 0.46 mmol),Intermediate 8 (120 mg, 0.304 mmol), triethylamine (64 μL, 0.46 mmol),and sodium triacetoxyborohydride (322 mg, 1.52 mmol) was stirred at roomtemperature for 2 days. The reaction mixture was diluted with DCM,washed with saturated NaHCO₃ solution, then brine, dried over anhydrousMgSO₄, filtered, and concentrated. Purification by preparative TLC(silica, 5% of 1/9 NH₄—OH/methanol in DCM) afforded two bandscorresponding to the cis isomer pair (top spot, 79.5 mg) and the transisomer pair (bottom spot, 49.6 mg).

Top spot: ESI-MS calc. for C23H28F6N2O2: 478; Found: 479 (M+H).

Bottom spot: ESI-MS calc. for C23H28F6N2O2: 478; Found: 479 (M+H).

EXAMPLE 162

Step A

A solution of Intermediate 3 (59 mg, 0.23 mmol), intermediate 16 (100.0mg, 0.231 mmol), diisopropylethylamine (40 μL, 0.23 mmol) and crushedmolecular sieves (4 Å, 50 mg) in dichloroethane (5 mL) was treated withsodium triacetoxyborohydride (245 mg, 1.16 mmol) and stirred at roomtemperature overnight. The reaction was quenched with saturated sodiumbicarbonate solution (10 mL) and diluted with an additional 10 mL ofDCE. The organic layer was separated and the aqueous washed withdichloromethane (2×5 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The residue was purified by preparative TLC (eluant: 0.5%NH₄OH: 5% MeOH: 94.5% CH₂Cl₂) to yield 135 mg (87%) of the product as amixture of four diastereomers. LC-MS calculated for C₃₀H₃₃ClF₆N₂O₄ is634.20, found (MH)⁺ 635.1 and (MH+2)⁺ 637.2.Step B

A solution of the product described in step A, example 162 (120 mg,0.190 mmol) in methanol (3 mL) was treated with a 0.5 M solution ofsodium methoxide in methanol (1 mL) and the resulting mixture stirredfor 1 h at room temperature. The mixture was evaporated under vacuum andthe residue purified by preparative TLC (eluant: 1.0% NH₄OH: 10% MeOH:89% CH₂Cl₂) to yield 95 mg (87%) of the final product as a mixture offour diastereomers. The single isomers were obtained by using a GilsonHPLC equipped with a Preparatory ChiralCel OD column eluting with 5%ethanol and 95% hexane with a flow rate of 9 ml/min. LC-MS calculatedfor C₂₃H₃₀F₆N₂O₃ is 496.22, found (MH)⁺ 497.3 for all 4 isomer.

EXAMPLE 163

A solution of intermediate 24 (13 mg, 0.093 mmol), intermediate 16 (37mg, 0.093 mmol), diisopropylethylamine (17 μL, 0.093 mmol) and crushedmolecular sieves (4 Å, 20 mg) in dichloroethane (3 mL) was treated withsodium triacetoxyborohydride (100 mg, 0.465 mmol) and stirred at roomtemperature overnight. The reaction was quenched with saturated sodiumbicarbonate solution (10 mL) and diluted with an additional 10 mL ofDCE. The organic layer was separated and the aqueous washed withdichloromethane (2×5 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The residue was purified by preparative TLC (eluant: 0.5%NH₄OH: 5% MeOH: 94.5% CH₂Cl₂) to yield 26 mg (63%) of the final productas a mixture of four diastereomers. LC-MS calculated for C₂₄H₃₂F₆N₂O₃ is510.24, found (MH)⁺ 511.2.

EXAMPLE 164

Step A

A solution of intermediate 3 (113 mg, 0.446 mmol), intermediate 14(200.0 mg, 0.446 mmol), diisopropylethylamine (78 μL, 0.45 mmol) andcrushed molecular sieves (4 Å, 100 mg) in dichloroethane (25 mL) wastreated with sodium triacetoxyborohydride (473 mg, 2.23 mmol) andstirred at room temperature overnight. The reaction was quenched withsaturated sodium bicarbonate solution (20 mL) and diluted with anadditional 20 mL of DCE. The organic layer was separated and the aqueouswashed with dichloromethane (2×10 mL). The organics were combined, driedover anhydrous sodium sulfate, filtered and evaporated under reducedpressure. The residue was purified by preparative TLC (eluant: 100%ethyl acetate) to yield 263 mg (98%) of the product as a mixture of fourdiastereomers. LC-MS calculated for C₃₀H₃₃ClF₆N₂O₅ is 650.26, found(MH)⁺ 651.2 and (MH+2)⁺ 653.3.Step B

A solution of the product described in step A, example 164 (263 mg,0.440 mmol) in methanol (5 mL) was treated with a 0.5 M solution ofsodium methoxide in methanol (1 mL) and the resulting mixture wasstirred for 1 h at room temperature. The mixture was evaporated undervacuum and the residue was purified by preparative TLC (eluant: 1.0%NH₄OH: 10% MeOH: 89% CH₂Cl₂) to yield 210 mg (86%) of the final productas a mixture of four diastereomers. The single isomers were obtained byusing a Gilson HPLC equipped with a Preparatory ChiralCel OD columneluting with 7% ethanol and 93% hexane with a flow rate of 9 ml/min.LC-MS calculated for C₂₃H₃₀F₆N₂O₄ is 512.22, found (MH)⁺ 513.3 for all 4isomer.

EXAMPLE 165

A solution of intermediate 24 (30 mg, 0.22 mmol), intermediate 14 (100mg, 0223 mmol), diisopropylethylamine (39 μL, 0.22 mmol) and crushedmolecular sieves (4 Å, 80 mg) in dichloroethane (15 mL) was treated withsodium triacetoxyborohydride (236 mg, 1.12 mmol) and stirred at roomtemperature overnight. The reaction was quenched with a saturated sodiumbicarbonate solution (15 mL) and diluted with an additional 10 mL ofDCE. The organic layer was separated and the aqueous layer was washedwith dichloromethane (2×5 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The residue was purified by preparative TLC (eluant: 0.5%NH₄OH: 5% MeOH: 94.5% CH₂Cl₂) to yield 84 mg (72%) of the final productas a mixture of four diastereomers. A second purification of thismaterial by preparative TLC (eluant 7% MeOH: 93% CH₂Cl₂), developedtwice, provided two bands which were assigned as less polar and morepolar and were presumed to be a mixture of two isomers each. LC-MScalculated for C₂₄H₃₂F₆N₂O₄ is 526.24, found (MH)⁺ 527.3 for both.

EXAMPLE 166

Step A

To a cooled (0° C.) solution of oxalyl chloride (237 μL, 2.72 mmol) andDMSO (386 μL, 5.44 mmol) in dichloromethane (15 mL), under nitrogen, wasadded dropwise a prepared solution of the product from step A,intermediate 24 (220 mg, 1.36 mmol) in DCM (10 mL) via syringe. Themixture was stirred for 30 minutes at 0° C. and then triethylamine (1.52mL, 10.9 mmol) was added via syringe and the resulting mixture wasstirred overnight allowing to warm to room temperature. The solution wasevaporated in vacuo and the residue was purified by preparative TLC(eluant: 60% ethyl acetate: 40% hexane) to afford the product (133 mg,66%) as a yellow oil.Step B

To a cooled (0° C.) solution of the product from step A, Example 166 (50mg, 0.31 mmol) in ether (3 mL), under nitrogen, was added dropwisemethylmagnesium chloride (208 μL, 0.625 mmol) via syringe and theresulting mixture was stirred at 0° C. for 3 h. The reaction wasquenched by the slow addition of a saturated solution of ammoniumchloride (2 mL) and then the organic layer was separated. The aqueouslayer was extracted with ether (3×5 mL) and the organics were combined,dried over anhydrous sodium sulfate, filtered, and evaporated undervacuum. The material was used without further purification for the nextreaction.

Yield was quantitative. ¹H NMR (CDCl₃, 500 MHz): 3.78-3.74 (m, 1H),3.45-3.38 (m, 2H), 3.42 (overlapped s, 3H), 3.31 (s, 3H), 3.26 (d, J=11Hz, 1H), 1.93 (ddd, J=2.7, 5.3, 14.8 Hz, 1H), 1.72 (ddd, J=4.8, 11.9,14.8 Hz, 1H), 1.34 (s, 3H).Step C

A solution of product from step B, Example 166 (50 mg, 0.31 mmol) inTHF/water (1 mL/0.1 mL) was treated with concentrated hydrochloric acid(0.1 mL) and the resulting solution was stirred at room temperature for1 h. The mixture was concentrated in vacuo to remove the THF and theaqueous layer was extracted with ether (6×5 mL). The organics werecombined, dried over anhydrous sodium sulfate, filtered, and evaporatedunder reduced pressure to afford the product (7.5 mg, 18%) as a clearfilm. ¹H NMR (CDCl₃, 500 MHz): 4.31 (ddd, J=3.2, 7.6, 11.2 Hz, 1H), 3.96(dd, J=1.7, 11.0 Hz, H), 3.65 (ddd, J=3.0, 12.1, 14.0 Hz, 1H), 3.36 (dd,J=1.5, 11.2 Hz, 1H), 2.91 (ddd, J=7.6, 11.9, 14.2 Hz, 1H), 2.50 (ddd,J=1.6, 2.9, 14.2 Hz, 1H), 1.51 (s, 3H).Step D

A solution of product described in step C, Example 166 (5 mg, 0.04mmol), intermediate 14 (17 mg, 0039 mmol), diisopropylethylamine (7 μL,0.039 mmol) and crushed molecular sieves (4 Å, 80 mg) in dichloroethane(2 mL) was treated with sodium triacetoxyborohydride (42 mg, 0.20 mmol)and stirred at room temperature overnight. The reaction was quenchedwith a saturated sodium bicarbonate solution (5 mL) and diluted with anadditional 5 mL of DCE. The organic layer was separated and the aqueouslayer was washed with dichloromethane (2×5 mL). The organics werecombined, dried over anhydrous sodium sulfate, filtered and evaporatedunder reduced pressure. The residue was purified by preparative TLC(eluant: 0.5% NH₄OH: 5% MeOH: 94.5% CH₂Cl₂) to yield 8.8 mg (43%) of thefinal product as a mixture of four diastereomers. LC-MS calculated forC₂₄H₃₂F₆N₂O₄ is 526.24, found (MH)⁺ 527.3.

EXAMPLE 167

Step A

This intermediate was prepared in an analogous fashion to that ofIntermediate 10, except iodomethane was used in place of 2-iodopropane.The cis and trans isomers were not separated; therefore, the compoundwas used as a mixture of two diastereomers. ¹H NMR (CDCl₃, 500 MHz):4.56 (br s, 1H), 4.18-4.10 (m, 1H) 4.03 (br s, 1H), 3.78-3.74 (m, 1H),2.60 (dd, J=7.8, 13.3 Hz, 1H), 2.20-2.00 (m,2H), 1.62-1.55 (m, 1H), 1.43(s, 9H), 1.31 (s, 3H).Step B

A mixture of the acid described in step A, Example 167 (500 mg, 2.06mmol), 3,5-bis(trifluoromethyl)benzylamine hydrochloride (575 mg, 2.06mmol), HOAt (280 mg, 2.06 mmol), N,N-diisopropylethylamine (356 μl, 2.06mmol) in dichloromethane (25 mL) was treated with1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (786 mg,4.10 mmol) and stirred at room temperature overnight. The reactionmixture was diluted with dichloromethane (30 mL), washed with water(2×20 mL), brine (1×30 mL), dried over anhydrous sodium sulfate and thesolvent was evaporated. The pure compound was obtained by purificationwith preparative TLC (eluant 30% ethyl acetate/70% hexane), 610 mg(64%). LC-MS calculated for C₂₁H₂₆F₆N₂O₃ is 468.18, found (MH)⁺ 469.3.¹H NMR (CDCl₃, 500 MHz) δ 7.80 (s, 1H), 7.72 (s, 2H), 6.18 (br s, 1H),4.60 (dd, J=6.1, 15.6 Hz, 1H), 4.52 (dd, J=6.0, 15.9 Hz, 1H), 4.03-4.00(m, 1H), 2.62 (dd, J=7.8, 13.6 Hz, 1H), 2.22-2.05 (m, 3H), 1.60-1.54 (m,1H), 1.45 (s, 9H), 1.39 (s, 3H).Step C

The product described in step B, Example 167 (610 mg, 1.30 mmol) wasdissolved with 4 N HCl in dioxane (8 ml) and the resulting solution wasstirred at room temperature for 1 h. The reaction was evaporated undervacuum to afford the product (484 mg, 92%) as a white powder. LC-MScalculated for C₁₆H₁₈F₆N₂O is 368.18, found (M+H)⁺ 369.3.Step D

A solution of product described in step C, Example 167 (100 mg, 0.250mmol), tetrahydro-4H-pyran-4-one (25 μL, 0.25 mmol),diisopropylethylamine (44 μL, 0.25 mmol) and crushed molecular sieves (4Å, 50 mg) in dichloroethane (10 mL) was treated with sodiumtriacetoxyborohydride (265 mg, 1.25 mmol) and stirred at roomtemperature overnight. The reaction was quenched with a saturated sodiumbicarbonate solution (15 mL) and diluted with an additional 10 mL ofDCE. The organic layer was separated and the aqueous washed withdichloromethane (2×10 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The residue was purified by preparative TLC (eluant: 0.5%NH₄OH: 5% MeOH: 94.5% CH₂Cl₂) to yield 105 mg (86%) of the final product(Example 271) as a mixture of two diastereomers. LC-MS calculated forC₂₁H₂₆F₆N₂O₂ is 452.19, found (M+H)⁺ 453.2.

EXAMPLE 168

Step A

This intermediate was prepared in an analogous fashion to that ofIntermediate 10, except iodoethane was used in place of 2-iodopropane.The cis and trans isomers were not separated; therefore, the compoundwas used as a mixture of two diastereomers. ¹H NMR (CDCl₃, 500 MHz):4.51 (br s, 1H), 3.98 (br s, 1H), 2.63 (dd, J=7.3, 13.0 Hz, 1H), 2.14(ddd, J=6.8, 7.3, 13.0 Hz, 1H), 2.08-2.00 (m,2H), 1.76-1.59 (m, 4H),1.44 (s, 9H), 1.31 (s, 3H), 1.28-1.23 (m, 2H), 0.83 (t, J=7.4 Hz, 3H).Step B

A mixture of the acid described in step A, Example 168 (528 mg, 2.06mmol), 3,5-bis(trifluoromethyl)benzylamine hydrochloride (575 mg, 2.06mmol), HOAt (280 mg, 2.06 mmol), N,N-diisopropylethylamine (356 μl, 2.06mmol) in dichloromethane (25 mL) was treated with1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (786 mg,4.10 mmol) and stirred at room temperature overnight. The reactionmixture was diluted with dichloromethane (30 mL), washed with water(2×20 mL), brine (1×30 mL), dried over anhydrous sodium sulfate and thesolvent was evaporated. The pure compound was obtained by purificationwith preparative TLC (eluant 25% ethyl acetate/75% hexane), 380 mg(38%). LC-MS calculated for C₂₂H₂₈F₆N₂O₃ is 482.21, found (M+H)⁺ 483.2.Step C

The product described in step B, Example 168 (380 mg, 0.79 mmol) wasdissolved with 4 N HCl in dioxane (8 ml) and the resulting solution wasstirred at room temperature for 1 h. The reaction was evaporated undervacuum to afford the product (321.4 mg, 97%) as a white powder. LC-MScalculated for C₁₇H₂₀F₆N₂O is 382.20, found (M+H)⁺ 383.2.Step D

A solution of the product described in step C, Example 272 (103 mg,0.250 mmol), tetrahydro-4H-pyran-4-one (25 mL, 0.25 mmol),diisopropylethylamine (44 μL, 0.25 mmol) and crushed molecular sieves (4Å, 50 mg) in dichloroethane (10 mL) was treated with sodiumtriacetoxyborohydride (265 mg, 1.25 mmol) and stirred at roomtemperature overnight. The reaction was quenched with a saturated sodiumbicarbonate solution (15 mL) and diluted with an additional 10 mL ofDCE. The organic layer was separated and the aqueous layer was washedwith dichloromethane (2×10 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The residue was purified by preparative TLC (eluant: 0.5%NH₄OH: 5% MeOH: 94.5% CH₂Cl₂) to yield 114 mg (91%) of the final product(Example 272) as a mixture of two diastereomers. LC-MS calculated forC₂₂H₂₈F₆N₂O₂ is 466.19, found (MH)⁺ 467.3.

EXAMPLE 169

Step A

To a solution of intermediate 20 (10 mg, 0.020 mmol) and DIEA (7.2 μL,0.040 mmol) in dichloromethane (2 mL) under nitrogen was addedbenzylchloroformate (3.8 μL, 0.026 mmol) and the resulting solution wasstirred at room temperature overnight. The mixture was evaporated underreduced pressure and the residue purified by preparative TLC (eluant:15% ethyl acetate/85% hexane) to afforded the product (6.3 mg, 52%) as ayellow film. LC-MS calculated for C₂₇H₂₉F₆N₃O₅ is 589.2, found (M+H)⁺590.4.Step B

The product described in step A, Example 169 (6 mg, 0.01 mmol) wasdissolved with 4 N HCl in dioxane (1 ml) and the resulting solution wasstirred at room temperature for 1 h. The reaction was evaporated undervacuum to afford the product (5.88 mg, 97%) as a yellow powder. LC-MScalculated for C₂₇H₂₉F₆N₃O₅ is 489.2, found (MH)⁺ 490.2.Step C

A solution of the product described in step B, Example 169 (5.9 mg,0.0090 mmol), tetrahydro-4H-pyran-4-one (3 μL, 0.01 mmol),diisopropylethylamine (2 μL, 0.01 mmol) and crushed molecular sieves (4Å, 5 mg) in dichloroethane (2 mL) was treated with sodiumtriacetoxyborohydride (10 mg, 0.05 mmol) and stirred at room temperatureovernight. The reaction was quenched with a saturated sodium bicarbonatesolution (5 mL) and diluted with an additional 5 mL of DCE. The organiclayer was separated and the aqueous layer was washed withdichloromethane (2×5 mL). The organics were combined, dried overanhydrous sodium sulfate, filtered and evaporated under reducedpressure. The residue was purified by preparative TLC (eluant: 0.5%NH₄OH: 5% MeOH: 94.5% CH₂Cl₂) to yield 4.48 mg (87%) of the finalproduct. LC-MS calculated for C₂₇H₂₉F₆N₃O₄ is 573.21, found (M+H)⁺574.2.

EXAMPLE 170

To a solution of the product described as Example 169 (2.1 mg, 0.0030mmol) and 1 equivalent of concentrated HCl in ethanol (1 mL) was added10% palladium on carbon (5 mg) and the resulting suspension was setunder a hydrogen atmosphere introduced via a 5 L balloon of hydrogengas. The mixture was stirred at room temperature for 2 h, then thehydrogen gas was evacuated and replaced with nitrogen. The mixture wasfiltered through a Gilman PTFE 0.45 μM syringe filter to remove thecatalyst. The filtrate was evaporated to afford 0.83 mg (52%) of thefinal product (example 276) as a white powder. LC-MS calculated forC₂₀H₂₅F₆N₃O₂ is 453.21, found (M+H)⁺ 454.3.

EXAMPLE 171

A solution of the ketone Intermediate 18 (200 mg, 1.0 mmol), amineIntermediate 14 (as a hydrochloride, 230 mg, 0.5 mmol),diisopropylethylamine (200 mg, 1.5 mmol), 4 Å molecular sieves (500 mg)in 10 mL of dry dichloroethane was treated with sodiumtriacetoxyborohydride (840 mg, 2.0 mmol) and the reaction mixture wasstirred at room temperature 16 h. The crude reaction mixture was pouredonto a saturated solution of sodium bicarbonate (50 mL), and the organicsolvent was slowly evaporated under mildly reduced pressure (250 torr)and slight heating (40° C.). HPLC analysis of the aqueous phaseindicated complete breakdown of the initial borane adduct afterapproximately 30 minutes. The crude product was extracted intodichloromethane, dried (anhydrous magnesium sulfate) and the solvent wasremoved in vacuo. The residue was further purified by preparative TLC toafford 50 mg (46%) of the higher eluting cis-isomeric mixture and 41 mgof the corresponding lower eluting trans-isomeric pair. The cis-isomericpair was separated into single diastereomers by chiral semi-preparativeHPLC using a ChiralPak OD column, eluted by a mixture of hexanes andethyl alcohol (95:5) at 9 mL/min. The retention time of the isomersunder the corresponding analytical conditions (1.0 mL/min flow rate)were 8.56 and 8.85 minutes, for the biologically less and more activeisomer, respectively. LC MS for C24H29F9N2O3 for [M+H]⁺ calc. 565, found565.

EXAMPLE 172

This compound, as a mixture of 4 isomers, was prepared starting fromIntermediate 14 and the less polar endo hydroxyketone (Intermediate 21)according to the procedure described under Example 171. LC MS forC26H34F6N2O4 for [M+H]⁺ calc. 523, found 523.

EXAMPLE 173

This compound (800 mg), as a mixture of 4 isomers, was prepared startingfrom the unsaturated ketone (850 mg) in step A of the Intermediate 22and Intermediate 14 (900 mg) according to the same procedure describedunder Example 171. LC MS for C26H32F6N2O3 for [M+H]⁺ calc. 535, found535.

EXAMPLE 174

A solution of the unsaturated ketone (1.38 g, 10.0 mmol) in step a ofIntermediate 22, amine Intermediate 14 (as a hydrochloride, 1.4 g mg,3.0 mmol), diisopropylethylamine (0.52 g, 4 mmol), 4 Å molecular sieves(1.0 mg) in 20 mL of dry dichloroethane was treated with sodiumtriacetoxyborohydride (2.10 g, 10 mmol) and the reaction mixture wasstirred at room temperature 48 h. 1.0 mL of a 30% aqueous formalinsolution was added and followed by 1.0 g of 4 Å molecular sieves and2.12 g of sodium triacetoxyborohydride. The mixture was stirred for 5 h.The crude reaction mixture was poured onto a saturated solution ofsodium bicarbonate (50 mL), and the organic solvent was slowlyevaporated under mildly reduced pressure (250 torr) and slight heating(60° C.). HPLC analysis of the aqueous phase indicated completebreakdown of the initial borane adduct after approximately 120 minutes.The crude product was extracted into dichloromethane, dried (anhydrousmagnesium sulfate) and the solvent was removed in vacuo. The residue wasfurther purified by preparative TLC to afford the desired endo and exomixture as a gummy solid (2.0 g). LC MS for C27H34F6N2O3 for [M+H]⁺calc. 549, found 549. Small quantities of (˜150 mg) endo (less polar)and exo (more polar) isomers were separated on preparative TLC.

EXAMPLE 175

Procedure A

A mixture of the Example 172 (800 mg) and 10% Pd/C (200 mg) in 25 mL ofmethanol was shaken on a Parr shaker for 2 h under 45 psi of hydrogen.The catalyst was removed by filtration and the filtrate was evaporatedto yield the desired product as a mixture of endo and exo isomers (790mg). The mixture was separated into single isomers by chiralsemi-preparative HPLC using a ChiralPak OD column, eluted by a mixtureof hexanes and ethyl alcohol (95:5) at 9 mL/min. The retention time ofthe isomers under the corresponding analytical conditions (1.0 mL/minflow rate) were 12.48 and 14.65 minutes, for the biologically more andless active isomer, respectively. LC MS for C26H34F6N2O3 for [M+H]⁺calc. 537, found 537.

Procedure B

The mixture of endo and exo isomers was also prepared starting from theIntermediate 22 and the Intermediate 14 according to the same proceduredescribed under Example 171. LC MS for C26H34F6N2O3 for [M+H]⁺ calc.537, found 537.

EXAMPLE 176

A mixture of the Example 174 (1.70 g) and 10% Pd/C (0.5 g) in 25 mL ofmethanol was shaken on a Parr shaker for 3 h under 50 psi of hydrogen.The catalyst was removed by filtration and the filtrate was evaporatedto yield the desired product as a mixture of endo and exo isomers (1.30g). The mixture was separated into single isomers by chiralsemi-preparative HPLC using a ChiralPak OD column, eluted by a mixtureof hexanes and ethyl alcohol (95:5) at 9 ml/min. The retention time ofthe isomers under the corresponding analytical conditions (1.0 mL/minflow rate) were 9.83 and 10.41 minutes, for the biologically more andless active isomer, respectively. LC MS for C27H36F6N2O3 for [M+H]⁺calc. 551, found 551.

EXAMPLE 177

A solution of Intermediate 23 (50 mg, 0.4 mmol), amine Intermediate 16(as a hydrochloride, 90 mg, 0.2 mmol), diisopropylethylamine (0.60 mg,0.4 mmol), 4 Å molecular sieves (50 mg) in 5 mL of dry dichloroethanewas treated with sodium triacetoxyborohydride (210 mg, 1.0 mmol) and thereaction mixture was stirred at room temperature 16 h. The crudereaction mixture was poured onto a saturated solution of sodiumbicarbonate (10 mL). The crude product was extracted intodichloromethane, dried (anhydrous magnesium sulfate) and the solvent wasremoved in vacuo. The residue was further purified by preparative TLC toafford the desired product as an oil (75 mg). LC MS for C24H31F7N2O2 for[M+H]⁺ calc. 513, found 513. The four single diastereoisomers wereobtained after twice chiral semi-preparative HPLC using a ChiralPak ODcolumn (first run: 6.15, 7.52 minutes) and AD column (second run: 5.98,6.92, 7.25, 8.32 minutes), eluted by a mixture of hexanes and ethylalcohol (90:10) at 9 mL/min.

EXAMPLE 178

Step A

To a solution of 1.5 g (15 mmol) of diisopropylamine in 50 mL of THF at−78° C. under nitrogen was added a solution of n-butyllithium (2.5 M,6.0 mL, 15 mmol) in hexane dropwise. The resulting mixture was warmed to0° C. for 30 minutes and then recooled at −78° C. A solution of 2.3 g(10 mmol) of tert-butyl 3,3-dimethoxy-cyclopetanecarboxylate in 10 mL ofTHP was added dropwise. The resulting red solution was stirred at −78°C. for 30 minutes. A neat solution of 3.0 g (30 mmol) of cyclohexanonewas added dropwise. After stirred for additional 1 h, the reaction wasquenched with aqueous saturated ammonium chloride and extracted withether. The title compound (1.60 g, 57%) was obtained as a colorless oilafter flash chromatography purification (10% EtOAc/Hexane). ¹H NMR (400MHz, CDCl₃): 1.10 (m, 1H), 1.43 (ss, 9H), 1.60 (m, 8H), 1.78 (m, 5H),2.30 (m, 1H), 2.40 (m, 1H), 3.42 (s, 3H), 3.46 (s, 3H).Step B

1.6 g (5.7 mmol) of the above ester (step A, Example 178) was mixed with20 mL of 1:1 (v/v) TFA/CH₂Cl₂ and allowed to stand at room temperaturefor 30 minutes. The reaction was concentrated under reduced pressure anddried under high vacuum overnight. The title compound (1.50 g) wasobtained as a yellow solid. ¹H NMR (400 MHz, CDCl₃): 1.18 (m, 1H),1.40-1.80 (m, 9H), 2.38 (m, 3H), 2.48 (d, 1H), 2.80 (d, 1H).Step C

A mixture of 0.33 g (1.5 mmol) of1-(2′-hydroxy-cyclohexyl)-3-oxo-cyclopentanecarboxylic acid (Step B,Example 178), 0.43 g (1.5 mmol) of (3,5-bis-trifluoromethyl)benzylaminehydrochloride, 0.23 g (1.6 mmol) of HOAt, 0.43 g (2.3 mmol) of EDC and0.2 g (1.5 mmol) of DIEA in 20 mL of CH₂Cl₂ was stirred for 2 h, dilutedwith CH₂Cl₂, washed with water, 2 N aqueous HCl and brine, dried overNa₂SO₄, evaporated and dried under vacuum. The title compound (0.36 g)was obtained as a light yellow solid after being purified on preparativeTLC (5% MeOH/CH₂Cl₂. ¹H NMR (400 MHz, CDCl₃): 1.18 (m, 1H), 1.35 (m,1H), 1.50 (m, 2H), 1.70 (m, 6H), 2.30 (m, 4H), 2.42 (d, 1H), 2.90 (d,1H), 4.60 (m, 2H), 7.74 (s, 2H), 7.78 (s, 1H), 7.85 (broad, 1H). LC MSfor C21H23F6NO3 for [M+H]⁺ calc. 452, found 452.Step D

A mixture of 0.35 g (0.78 mmol) of,N-(3,5-bis-trifluoromethyl-benzyl)-1-(2′-hydroxy-cyclohexyl)-3-oxo-cyclopentanecarbamide(Step C, Example 178), 0.3 g (2 mmol) of tetrahydro-4H-pyran-4-ylammonium chloride (Intermediate 2), 0.26 g (2.0 mmol) of DIEA, 0.4 g (2mmol) of sodium triacetoxyborohydride and 0.5 g of molecular sieves (4Å) in 10 mL of CH₂Cl₂ was stirred overnight and then quenched with 20 mLof saturated aqueous Na₂CO₃. The solid was removed by filtration andwashing with CH₂Cl₂. CH₂Cl₂ was removed and the mixture was heated at60° C. for 30 minutes. After being cooled, the aqueous solution wasextracted with CH₂Cl₂ (3×) and the organic phases were dried withNa₂SO₄, and evaporated. The residue was purified by preparative TLC (10%[aq. NH₄OH/MeOH 1/9]/CH₂Cl₂). Two components were obtained. Less polar(0.13 g, cis isomer) ¹H NMR (400 MHz, CDCl₃); 1.00-2.05 (m, 20H), 2.30(m, 1H), 2.60 (m, 1H), 3.30 (m, 2H), 3.55 (broad, 1H), 3.95 (m, 2H),4.48 (m, 2H), 5.40 (s, 1H), 7.71 (s, 2H), 7.78 (s, 1H), 9.95 (broad,1H). LC MS for C26H34F6N2O3 for [M+H]⁺ calc. 537, found 537. More polar(0.09 g, trans isomer) ¹H NMR (400 MHz, CDCl₃): 1.00-2.05 (m, 20H), 2.42(m, 1H), 2.70 (m, 1H), 3.30 (m, 2H), 3.55 (broad, 1H), 3.90 (m, 2H),4.50 (m, 2H), 5.40 (s, 1H), 7.71 (s, 2H), 7.78 (s, 1H), 9.95 (broad,1H). LC MS for C26H34F6N2O3 for [M+H]⁺ calc. 537, found 537.

EXAMPLE 179

This compound, as a mixture of cis and trans isomers, was preparedstarting from cyclopentanone according to the same procedure asdescribed under Example 178. The cis (less polar) and trans (more polar)isomers were separated on preparative TLC. The cis isomer was furtherseparated into single isomers by chiral semi-preparative HPLC using aChiralPak OD column, eluted by a mixture of hexanes and ethyl alcohol(95:5) at 9 mL/min. The retention time of the isomers under thecorresponding analytical conditions (1.0 mL/min flow rate) were 14.71and 16.39 minutes, for the biologically less and more active isomer,respectively. LC MS for C25H32F6N2O3 for [M+H]⁺ calc. 523, found 523.

EXAMPLE 180

This compound, as a mixture of cis and trans isomers, was preparedstarting from cyclobutanone according to the same procedure describedunder Example 178. The cis (less polar) and trans (more polar) isomerswere separated on preparative TLC. The cis isomer was further separatedinto single isomers by chiral semi-preparative HPLC using a ChiralPak ODcolumn, eluted by a mixture of hexanes and ethyl alcohol (95:5) at 9ml/min. The retention time of the isomers under the correspondinganalytical conditions (1.0 mL/min flow rate) were 15.18 and 25.46minutes, for the biologically less and more active isomer, respectively.LC MS for C24H30F6N2O3 for [M+H]⁺ calc. 509, found 509.

EXAMPLE 181

Step A

To a flame-dried 1000 mL round-bottomed flask, was added dry THF (150mL). The solvent was cooled to −78° C. before diisopropylamine (8.04 mL,57.4 mmol), 2.5 M n-butyllithium (22.95 mL, 57.37 mmol), and a solutionof the Schiff base prepared in Step C, Intermediate 9 (20 g, 52 mmol) indry THF (100 mL), were added sequentially. The reaction mixture wasstirred at −78° C. for 0.5 h before cyclopentanone (13.84 mL, 156.5mmol) was slowly added. After the reaction was stirred for another 2 h,the mixture was quenched with a saturated NH₄Cl solution, extracted withether (three times), washed by brine, dried over Na₂SO₄, andconcentrated. The crude product was purified by MPLC (15% EtOAc/Hexanes)to yield the desired product (2.35 g, 9.6%). ¹H-NMR (400 MHz, CDCl₃):7.1-7.6 (m, 15H), 5.23 (s, 2H), 3.80 (m, 2H), 2.60 (m, 1H), 2.40 (m,1H), 1.50-2.20 (m, 8H). LC-MS calc. For C31H33NO3: 467.25; Found: 468(M+H).Step B

To a solution of the intermediate prepared in Step A, Example 181 (2.35g, 5.03 mmol) in THP (20 mL), was slowly added 2 N HCl (8 mL, 0.02 mol)to form a homogeneous solution. The reaction mixture was stirred at roomtemperature overnight, and concentrated under vacuum. The residualsolution was extracted with hexanes (three times) and the aqueoussolution was diluted by water (80 mL), and made alkaline by the additionof solid NaHCO₃ (3.4 g, 40 mmol). The resulting product was dissolved inDCM (150 mL) and di-tert-butyl dicarbonate (3.29 g, 15.1 mmol) wasadded. The reaction was then stirred at room temperature for 24 h. Theaqueous phase was separated and extracted by DCM (twice). The combinedorganic portion was washed with water and brine, dried over Na₂SO₄,filtered and concentrated. The residue was purified by flash columnchromatography (silica gel, 20% EA/hexanes to 30% EA/hexanes) to yieldthe desired intermediate (1.03 g, 50.7%). ¹H-NMR (500 MHz, CDCl₃)7.36-7.42 (m, 5H), 5.20 (d, 2H), 4.67 (s, 1H), 4.27 (s, 1H), 3.17 (s,1H), 2.18 (m, 2H), 2.05 (m, 2H), 1.82 (m, 2H), 1.45-1.64 (m, 8H), 1.45(s, 9H).Step C

To a solution of the intermediate prepared in Step B, Example 181 (1.0g, 2.5 mmol) in EtOAc (100 mL), was added 10% Pd/C (75 mg). The reactionmixture was placed in a Parr-shaker under 50 psi of H₂ for 8 h. Thesolution was filtered through celite and concentrated under vacuum toyield the desired intermediate (780 mg, 100%). ¹H-NMR (400 MHz, CDCl₃):4.10 (br s, 1H), 5.20 (d, 2H), 2.25 (m, 2H), 2.00 (m, 2H), 1.65-1.75 (m,9H), 1.45 (s, 9H).Step D

To a flask was added the intermediate prepared in Step C, Example 181(400 mg, 1.28 mmol), 3,5-bis(trifluoromethyl)benzylamine HCl salt (713mg, 2.55 mmol), EDC (488 mg, 2.55 mmol), HOAt (174 mg, 1.28 mmol) in DCM(10 mL). The reaction was stirred for 30 minutes and DIEA (444 μL, 2.55mmol) was added into mixture. The resulting mixture was then stirred for2 h. The reaction was diluted by DCM, washed by water, brine, dried overNa₂SO₄, filtered and concentrated. The residue was purified by flashcolumn chromatography (silica gel, 30% EtOAc/Hexane to 40% EtOAc/Hexane)to yield the BOC protected Intermediate (1.01 g, 56.7%). ¹H-NMR (500MHz, CDCl₃): 8.60 (br s, 1H), 7.78 (s, 1H), 7.75 (s, 2H), 5.24 (br d,1H), 4.58 (d, J=6.0 Hz, 2H), 3.96 (m, 1H), 3.18 (br s, 1H), 2.12-2.24(m, 2H), 1.90-1.98 (m, 2H), 1.56-1.84 (m, 10H), 1.41 (s, 9H). ESI-MScalc. For C23H32F3N3O3: 455.24; Found: 356 (M+H-100). The BOC protectedintermediate was treated with 4 N HCl/Dioxane to yield the desiredintermediate (450 mg).Step E

A mixture of the intermediate prepared in Step D, Example 181 (207 mg,0.436 mmol), Intermediate 5 (174 mg, 1.53 mmol), molecular sieve (4 Å,450 mg), DIEA (76 μL, 0.44 mmol) and sodium triacetoxyborohydride (323mg, 1.53 mmol in DCM (10 mL) was stirred for 24 h. The reaction mixturewas filtered, washing with DCM and MeOH, mixed with saturated aqueousNaHCO₃, and heated at 60° C. for 1.5 h before the DCM and methanol wereevaporated. The resulted mixture was diluted with water (10 mL) andextracted with DCM (five times). The organic portion was separated,washed with water and brine, dried over Na₂SO₄, filtered andconcentrated. The residue was purified on preparative TLC (1000 micron)(developed by 4% [aqueous NH₄OH/MeOH(1/9)] in DCM) to yield the finaltitle compound as a free base. Its HCl salt Example 181 (112 mg) wasformed by treatment with 4 N HCl/dioxane. ¹H-NMR (400 MHz, CDCl₃): 10.14(br s, 1H), 7.80 (s, 1H), 7.70 (s, 2H), 4.50 (m, 2H), 3.55-3.85 (m, 2H),3.30-3.42 (m, 2H), 2.75 (br s, 1H), 1.82-2.24 (m, 8H), 1.45-1.75 (m,12H), 0.92 (m, 3H). ESI-MS calc. for C26H34F6N2O3: 536.25; Found: 537(M+H).

EXAMPLE 182

This compound, as a mixture of cis and trans isomers, was prepared viareplacement of cyclohexanone with cyclohexyl iodide (5.0 eq.) in HMPA(1.0 eq.) (alkylation time: 12 h in Step A) according to the sameprocedure described under Example 178. The cis (less polar) and trans(more polar) isomers were separated on preparative TLC. LC MS forC26H34F6N2O2 for [M+H]⁺ calc. 521, found 521.

EXAMPLE 183

This compound, as a mixture of cis and trans isomers, was prepared viareplacement of cyclohexanone with cyclopentyl iodide (5.0 eq.) in HMPA(1.0 eq.) (alkylation time: 12 h in Step A) according to the sameprocedure described under Example 178. The cis (less polar) and trans(more polar) isomers were separated on preparative TLC. LC MS forC25H32F6N2O2 for [M+H]⁺ calc. 507, found 507.

EXAMPLE 184

This compound, as a mixture of cis and trans isomers, was prepared viareplacement of cyclohexanone with cyclobutyl bromide (5.0 eq.) in HMPA(1.0 eq.) (alkylation time: 12 h in Step A) according to the sameprocedure described under Example 178. The cis (less polar) and trans(more polar) isomers were separated on preparative TLC. LC MS forC24H30F6N2O2 for [M+H]⁺ calc. 493, found 493.

EXAMPLE 185

This compound, as a mixture of cis and trans isomers, was prepared viareplacement of cyclohexanone with N-Cbz-piperidyl bromide (5.0 eq.) inHMPA (1.0 eq.) (alkylation time: 12 h in Step A) according to the sameprocedure described under Example 178. The cis (High Band) and trans(Low Band) isomers were separated on preparative TLC. LC MS forC33H39F6N3O4 for [M+H⁺ calc. 656, found 656.

EXAMPLE 186

To a stirred mixture of Example 185 (high band, cis isomer, 660 mg, 1.0mmol) and 4 Å molecular sieves (2.0 g) in 20 mL of dichloromethane wasadded 0.5 mL of a 30% aqueous formalin solution. The mixture was stirredfor 5 minutes and sodium triacetoxyborohydride (420 mg, 2.0 mmol) wasadded. The mixture was stirred for 2 h, quenched with saturated aqueoussodium carbonate, filtered, and washed with dichloromethane. The crudeproduct was extracted into dichloromethane, dried over Na₂SO₄,evaporated and purified on preparative TLC. The desired product wasobtained as a gummy solid (520 mg, 78%). LC MS for C34H41F6N3O4 for[M+H]⁺ calc. 670, found 670.

EXAMPLE 187

A mixture of Example 186 (500 mg) and 10% Pd/C (100 mg) in 20 mL ofmethanol was hydrogenated on a Parr-shaker for 5 h under 45 psi ofhydrogen. The catalyst was removed by filtration. The filtrate wasevaporated. The desired product was obtained as a light yellow solid(400 mg, 100%). LC MS for C26H35F6N3O2 for [M+H]⁺ calc. 536, found 536.

EXAMPLE 188

A mixture of Example 187 (54 mg, 0.10 mmol), pyridine (0.2 mL) andacetic anhydride (0.1 mL) in 1 mL of dichloromethane was stirredovernight. Preparative TLC afforded the desired product as a white solid(32 mg, 55%). LC MS for C28H37F6N3O3 for [M+H]⁺ calc. 578, found 578.

EXAMPLE 189

A mixture of Example 187 (54 mg, 0.10 mmol), pyridine (0.2 mL) and mesylchloride (0.1 mL) in 1 mL of dichloromethane was stirred overnight.Preparative TLC afforded the desired product as a white solid (37 mg,60%). LC MS for C27H37F6N3O4S for [M+H]⁺ calc. 614, found 614.

EXAMPLE 190

This compound, as a mixture of cis and trans isomers, was prepared viareplacement of cyclohexanone with benzyl bromide (5.0 eq.) in HMPA (1.0eq.) (alkylation time: 12 h in Step A, Example 178) according to thesame procedure described under Example 178. LC MS for C₂₇H₃₀F₆N₂O₂ for[M+H]⁺ calc. 529, found 529.

EXAMPLE 191

This compound, as a mixture of cis and trans isomers, was preparedstarting from Example 190 according to the procedure described underExample 186. LC MS for C28H32F6N2O2 for [M+H]⁺ calc. 542, found 542.

EXAMPLE 192

This compound, as a mixture of all possible isomers, was preparedstarting from 2,2-dimethyltetrahydro-2-H-pyran-4-amine and Intermediate8 according to the same procedure described under Example 1. LC MS forC25H34F6N2O2 for [M+H]⁺ calc. 509, found 509.

EXAMPLE 193

Step A

To a solution of bis(trifluoromethyl)benzaldehyde (20 g, 0.083 mol) in200 mL of THF at −78 C was added dropwise a solution of 84 mL ofmethylmagnesium bromide (1 M, 0.08 mol) in butyl ether. The temperaturewas raised up to room temperature and the entire mixture was poured intoa stirred mixture of ammonium chloride, ice and water (1000 mL) andextracted with ethyl acetate (2×1000 mL). The organic phases were driedover Na₂SO₄. Evaporation in vacuo afforded the title compound as a lightyellow liquid (20.64 g, 98%), which was used directly for furtherconversion.Step B

To a stirred solution of bis-(trifluoromethyl)phenylethanol (20.6 g,80.0 mmol), phthalimide (11.8 g, 80.0 mmol) and triphenylphosphine (22.6g, 100 mmol) in 150 mL of THF at 0° C. was added dropwise a solution ofDEAD (17.4 g, 100 mmol) in 100 mL of THF over 30 minutes. The mixturewas then stirred at room temperature overnight and condensed in vacuo.Flash chromatography on silica gel (500 g) afforded the title compoundas a light yellow solid. ¹H NMR (400 MHz, CDCl₃): 1.96 (d, 3H), 5.64 (q,1H), 7.70 (m, 2H), 7.79 (s, 1H), 7.80 (m,2H), 7.96 (s, 2H).Step C

A mixture of N-(bis-[trifluoromethyl]phenylethyl)phthalimide (allmaterial, ˜0.076 mol) and hydrazine (3.2 g, 100 mmol) in 500 mL ofethanol was stirred at 80° C. for 2 h. The flask was put into therefrigerator overnight. The solid was removed by filtration and washingwith ethanol. The filtrates were combined and evaporated in vacuo toafford the crude product which was stirred with di-tert-butyldicarbonate (17 g, 80 mmol) in 200 mL of dioxane for 30 minutes and thenevaporated in vacuo. The residue was purified by flash chromatography onsilica gel (400 g) using 30% EtOAc/hexanes. The BOC-Amide (20.7 g) wasobtained as a white solid. ¹H NMR (400 MHz, CD₃OD): 1.40 (s, 9H), 1.71(d, 3H), 4.50 (m, 1H), 7.75 (s,3H). This material was stirred with asolution of 100 mL of 4 N HCl in dioxane for 2 h. The mixture wasevaporated and dried in vacuo to afford the title compound as a whitesolid (15.6 g). ¹H NMR (400 MHz, CD₃OD): 1.69 (d, 2H), 4.75 (q, 1H),8.05 (s, 1H), 8.16 (s, 2H).Step D

This compound, as a mixture of two diastereoisomers, was prepared fromIntermediate 9 and the amine prepared in Step C of Example 193,according to the same procedure as described under Step A, Example 20.LC MS for C24H32F6N2O4 for [M+H]⁺ calc. 527, found 527.Step E

This compound, as a mixture of two diastereoisomers, was preparedfollowing the procedure described under Step B, Example 20. LC MS forC19H24F6N2O2 for [M+H]⁺ calc. 427, found 427.Step F

This compound, as a mixture of two diastereoisomers, was preparedaccording to the procedure described under Step C, example 20. LC MS forC24H32F6N2O3 for [M+H⁺ calc. 511, found 511.

EXAMPLE 194

Step A

To a stirred, −78° C. solution of the Schiff base prepared in Step C,Intermediate 9 (38.4 g, 100 mmol) in 200 mL of THF was added a solutionof LDA (2.0 M, 55 mL, 110 mmol) in THF. The mixture was stirred for 30minutes at −78° C., then a solution of allyl bromide (20 mL, 200 mmol)in HMPA (18 mL, 100 mmol) was added dropwise. The resulting red solutionwas stirred at −78° C. for 1 h, warmed to room temperature by removingthe cooling bath, diluted with water, and extracted with ether. Theether layer was washed with water and brine, dried over Na₂SO₄ andevaporated. The residue was dissolved in 300 mL of THF. To this solutionwas added 150 mL of 2 N aqueous HCl and stirred for 1 h, evaporated toremove THF, and extracted with hexane (3×). The aqueous solution wasmade alkaline to pH>9 with a saturated aqueous sodium carbonate solutionand immediately was mixed and stirred with a solution of di-tert-butyldicarbonate (42 g, 200 mmol) in 200 mL of dichloromethane. After 30minutes, the organic phase was separated and the aqueous layer wasextracted with dichloromethane (2×). The combined organic phases werewashed with water and brine, dried over Na₂SO₄ and evaporated. Theresidue was purified on by flash chromatography (10% EtOAc/Hexane) toyield a mixture of cis and trans isomers (24.0 g, 65%). The mixture wasseparated into single cis (fast-eluted) and trans (slow-eluted) isomerson MPLC (5% EtOAc/Hexane). ¹H NMR (400 MHz, CDCl₃): cis: 7.40 (m, 5H),5.68 (m, 1H), 5.18 (s, 2H), 5.04 (m, 2H), 4.85 (br s, 1H), 4.10 (br s,1H), 2.50 (dd, J=7.2 Hz, 1H), 2.30 (dd, J=7.3 Hz, 1H), 2.20 (m, 1H),2.00 (m, 3H), 1.70-1.43 (m, 2H), 1.44 (s, 9H). trans: 7.38 (m, 5H), 5.65(m, 1H), 5.12 (s, 2H), 5.03 (m, 2H), 4.50 (br s, 1H), 4.00 (br s, 1H),2.62 (dd, J=6.1 Hz, 1H), 2.24 (m, 2H), 2.10 (m, 2H), 1.70 (m, 1H), 1.41(s, 9H), 1.42-1.30 (m, 2H).Step B

A mixture of the cis ester prepared in Step A, Example 194 (0.65 g) and10% Pd/C (0.2 g) in methanol (mL) was shaken on a Parr apparatus for 2 hunder 50 psi of hydrogen. The catalyst was removed by filtration, thefiltrate was evaporated and dried under vacuum to yield the desired acidas a white solid which was used in next step without furtherpurification.Step C

A mixture of the cis acid prepared in Step B, Example 194 (101 mg, 0.4mmol), the amine prepared in Intermediate 6, as a HCl salt (216 mg, 0.8mmol) and EDC (190 mg, 1.0 mmol) in 2 mL of dichloromethane was stirredat room temperature overnight., diluted with dichloromethane, washedwith water, 1 N aqueous HCl and brine, dried over Na₂SO₄, evaporated andpurified on preparative TLC (5% MeOH/Hexane). The desired product wasobtained as a gummy solid (100 mg, 51%). ¹H NMR (400 MHz, CDCl₃): 7.76(s, 111, 7.71 (s, 2H), 6.72 (br s, 1H), 5.29 (br s, 1H), 4.56 (d, J=6.1Hz, 2H), 4.02 (m, 1H), 2.10 (m, 2H), 2.00 (m, 1H), 1.80 (dd, J=3.0, 5.2Hz, 1H), 1.68 (m, 1H), 1.50 (m, 3H), 1.40 (s, 9H), 1.20 (m, 2H), 0.92(t, J=7.2 Hz, 3H).Step D

The BOC-Amide from Step C, Example 194 (100 mg, 0.2 mmol) was treatedwith 4 N HCl/dioxane (5 mL) for 5 h, evaporated and dried under vacuumto afford a white solid (86 mg, 99%). LC MS for C18H22F6N2O for [M+H]⁺calc. 397, found 397.Step E

The amino amide prepared in Step D, Example 194 (86 mg, 0.2 mmol),tetrahydro-4H-pyran-4-one (100 mg, 1.0 mmol), sodiumtriacetoxyborohydride (210 mg, 1.0 mmol), DIEA (130 mg, 1.0 mmol) and 4Å molecular sieves (100 mg) in 5 mL of dichloromethane was stirredovernight, quenched with saturated aqueous sodium carbonate andfiltered. The crude product was extracted into dichloromethane, driedover Na₂SO₄ and evaporated. Purification on preparative TLC (10% [aq.NH₄OH/MeOH 1/9]/DCM) gave the desired product as an oil (72 mg, 75%). LCMS for C23H30F6N2O2 for [M+H]⁺ calc. 481, found 481.

EXAMPLE 195

Step A

A neat mixture of 54 g (0.29 mole) ethyl (2-aminothiazol-4-yl)acetateand 50 g (0.28 mole) benzophenone imine was stirred at 190° C. for 5 hand then cooled to room temperature and diluted with 100 mL of CH₂Cl₂.The entire mixture was transferred onto a silica gel column and elutedwith 20% EtOAc/Hexane. The title compound was obtained as light-yellowsolid (70 g, 69% yield). ¹H NMR (300 MHz, CDCl₃): 1.26 (t, 3H), 3.74 (s,2H), 4.15 (q, 2H), 6.87 (s, 1H), 77.25-7.86 (m, 10H). LC MS forC20H18N2O2S for [M+H]⁺ calc. 351, found 351.Step B

To a mixture of 35 g (0.10 mol) of ethyl(2-diphenylmethyleneamino-thiazol-4-yl)acetate (Step A, Example 195),cis-1,3-dichloro-2-butene (13 mL, 0.11 mol) in 500 mL of DME at roomtemperature was added in multiple portions solid NaH (60% oil, 10 g,0.25 mol). The resulting mixture was stirred for 2 days, poured into2000 mL of ice-water and extracted with 1500 mL of ether. The etherlayer was washed with water (3×500 mL), dried over Na₂SO₄ andevaporated. Flash chromatography (silica gel, 5% EtOAc/Hexane) affordedthe title compound as an oil (24 g, 59%). ¹H NMR (300 MHz, CDCl₃): 1.20(t, 3H), 2.87 (d, 2H), 3.19 (d, 2H), 4.14 (q, 2H), 5.29 (s, 2H), 6.71(s, 1H), 7.26-7.81 (m, 10H). LC MS for C24H22N2O2S for [M+H]⁺ calc. 403,found 403.Step C

24 g (0.059 mol) of ethyl1-(2-diphenylmethyleneamino-thiazol-4-yl)-3-cyclopentenecarboxylate(Step B, Example 195) was dissolved in 100 mL of 4 N HCl/dioxane. After1 h, 1.8 mL of water was added. The mixture was stirred for 3 h andevaporated to dryness. The residue was dissolved in 100 mL of CH₂Cl₂ and15 mL of DIEA was added. The entire mixture was dumped onto a silica gelcolumn, eluted with 20% EtOAc/hexanes to remove benzophenone, theneluted with 40% EtOAc/hexane to give the title compound as a lightyellow solid (12.0 g, 85%). ¹H NMR (300 MHz, CDCl₃): 1.19 (t, 3H), 2.79(d, 12H), 3.15 (d, 2H), 4.13 (q, 2H), 5.66 (s, 2H), 5.82 (wide, 2H),6.19 (s, 1H).Step D

A mixture of 12 g (50 mmol) of ethyl1-(2-amino-thiazol-4-yl)-3-cyclopentenecarboxylate (Step C, Example195), 28 g (0.13 mol) of di-tert-butyl dicarbonate and 0.6 g of DMAP in250 mL of CH₂Cl₂ was stirred overnight, and evaporated. The titlecompound (21.0 g, 96%) was obtained as a yellow oil after flashchromatography purification on silica gel (10% EtOAc/Hexane). ¹H NMR(300 MHz, CDCl₃): 1.18 (t, 3H), 1.49 (d, 18H), 2.88 (d, 2H), 3.18 (d,2H), 4.13 (q, 2H), 5.65 (s, 2H), 6.83 (s, 1H). LC MS for C21H30N2O6S for[M+H]⁺ calc. 439, found 439.Step E

To a solution of 13 g (30 mmol) of ethyl1-(2-Bis-Boc-amino-thiazol-4-yl)-3-cyclopentenecarboxylate (Step D,Example 195) in 50 mL of anhydrous ether at −78° C. was added dropwise asolution of borane-dimethyl sulfide in THF (14 mL, 0.024 mmol). Thecooling bath was removed and the mixture was stirred at room temperaturefor 3 h, diluted with 250 mL of CH₂Cl₂, and 25 g of sodium acetate and55 g of PCC were added. The mixture was stirred overnight. The entiremixture was dumped onto a silica gel column and eluted with in 10%EtOAc/hexane and then 30% EtOAc/hexane. Two components were obtained.The fast-eluted isomer (yellow oil, 6.0 g) was identified as the titlecompound. ¹H NMR (300 MHz, CDCl₃): 1.21 (t, 3H), 1.50 (s, 18H), 2.33(t,2H), 2.42-2.70 (m, 2H), 2.78-3.10 (dd, 2H), 4.18 (q, 3H), 6.88 (s,1H). LC MS for C21H30N2O7S for [M+H]⁺ calc. 455, found 455.Step F

The slow-eluted component from the flash chromatography in Step E,Example 195 was proved to be the title compound (gummy material, 1.80g). ¹H NMR (300 MHz, CDCl₃): 1.16 (t, 3H), 1.46 (s, 9H), 2.27 (3, 2H),2.38-2.62 (m,2H), 2.64-3.00 (dd, 2H), 4.11 (q, 2H), 6.66 (s, 1H). LC MSfor C16H22N2O5S for [M+H]⁺ calc. 355, found 355.Step G

A mixture of 1.4 g (4.0 mmol) of ethyl1-(2-tert-butoxycarbonyl-amino-thiazol-4-yl)-3-oxo-cyclopentanecarboxylate(Step F, Example 195) and 0.82 g (13 mmol) of lithium hydroxidemonohydrate in a solution of 20 mL of MeOH and 2 mL of water was stirredat room temperature overnight. The entire mixture was poured onto asilica gel column and eluted with 10% MeOH/CH₂Cl₂. Evaporation in vacuoafforded a light yellow solid. 1.30 g of the title product was obtainedas a fluffy solid. ¹H NMR (300 MHz, CDCl₃): 1.52 (t, 9H), 2.10-3.20 (m,8H), 6.60 (s, 1H).Step H

A mixture of 0.65 g (2.0 mmol) of1-(2-tert-butoxycarbonyl-amino-thiazol-4-yl)-3-oxo-cyclopentanecarboxylic acid (Step H, Example 195), 0.70 g (2.5 mmol) of(3,5-bis-trifluoromethyl)benzylamine hydrochloride and 0.95 g EDC (5.0mmol) in 50 mL of CH₂Cl₂ was stirred for 2 h. The reaction mixture wasdiluted with 100 mL of CH₂Cl₂ and washed with 3 N aqueous HCl (3×50 mL),saturated aqueous NaHCO₃ (50 mL), and water (100 mL) and dried overNa₂SO₄ and evaporated in vacuo. 1.0 g of the title compound was obtainedas a yellow solid. ¹H NMR (400 MHz, CDCl₃): 1.55 (s, 9H), 2.10-2.22 (m,2H), 2.38-2.64 (m, 2H), 2.70-3.23 (dd,2H), 4.48-4.64 (m, 2H), 6.74 (s,1H), 7.36 (broad, 1H), 7.63 (s, 2H), 7.77 (s, 1H), 7.98 (broad, 1H). LCMS for C23H23F6N3O4S for [M+H]⁺ calc. 552, found 552.Step I

A mixture of 0.55 g (1.0 mmol) of,N-(3,5-bis-trifluoromethyl-benzyl)-1-(2-tert-butoxycarbonyl-amino-thiazol-4-yl)-3-oxo-cyclopentanecarbamide(Step H, Example 195), 0.27 g (2.0 mmol) of tetrahydro-4H-pyran-4-ylammonium chloride (Intermediate 2), 0.3 g (2 mmol) of DIEA, 0.83 g (4.0mmol) of sodium triacetoxyborohydride and 3.0 g of molecular sieves (4Å) in 50 mL of CH₂Cl₂ was stirred overnight and quenched with 50 mL ofsaturated aqueous Na₂CO₃. The solid was removed by filtration andwashing with CH₂Cl₂. The organic phase was separated and dried withNa₂SO₄ and evaporated. The residue was purified by preparative TLC (10%[aq. NH₄OH/MeOH 1/9]/CH₂Cl₂). The title compound (0.52 g, 82%) wasobtained as a mixture of cis and trans isomers. ¹H NMR (400 MHz, CDCl₃):1.30-2.60 (m, 25H), 3.97(m, 2H), 4.50 (m, 2H), 6.72 (s, 1H), 6.93(broad, 1H), 7.30 (broad, 1H), 7.58 (d, 2H), 7.75 (s,1H). LC MS forC₂₈H₃₄F₆N₄O₄S for [M+H]⁺ calc. 637, found 637.

EXAMPLE 196

A mixture of 0.32 g (0.50 mmol) ofN-(3,5-bis-trifluoromethyl-benzyl)-1-(2-tert-butoxycarbonyl-amino-thiazol-4-yl)-3-(tetrahydro-4H-pyran-4-ylamino]-cyclopentane-carbamide(Example 195), 0.3 g of a 37% aqueous formalin solution and 3.0 g ofmolecular sieves (4 Å) in 50 mL of CH₂Cl₂ was stirred for 30 minutes,then 0.84 g of sodium triacetoxyborohydride was added. The mixture wasstirred overnight and quenched with 50 mL of saturated aqueous Na₂CO₃.The solid was removed by filtration and washing with CH₂Cl₂. The organicphase was separated and dried with Na₂SO₄ and evaporated. The residuewas purified by preparative TLC (10% [aq. NH₄OH/MeOH 1/9]/CH₂Cl₂). Thetitle compound (0.250 g, 77%) was obtained as a mixture of cis and transisomers. ¹H NMR (400 MHz, CDCl₃): 1.20-3.49 (m, 28H), 4.00(m, 2H), 4.50(m, 2H), 6.70, 6.73 (ss, 1H), 6.93, 7.12 (broad, broad, 1H), 7.57 (d,2H), 7.78 (s, 1H), 8.27 (broad,1H). LC MS for C29H36F6N4O4S for [M+H]⁺calc. 651, found 651.

EXAMPLE 197

A mixture of 0.22 g (0.34 mmol) ofN-(3,5-bis-trifluoromethyl-benzyl)-1-(2-tert-butoxycarbonyl-amino-thiazol-4-yl)-3-(N-methyl,N-tetrahydro-4H-pyran-4-ylamino]-cyclopentanecarbamide (Example 196) and5.0 mL of TFA was allowed to stand at room temperature for 30 minutes,evaporated and dried in vacuo. The residue was dissolved in 5 mL of 4 NHCl in dioxane, evaporated and dried in vacuo. The hydrochloride salt ofthe title compound (mixture of cis and trans isomers, 210 mg, 100%) wasobtained as a light brown solid. LC MS for C24H28F6N4O2S for [M+H]⁺calc. 551, found 551.

EXAMPLE 198

A mixture of 0.055 g (0.10 mmol) ofN-(3,5-bis-trifluoromethyl-benzyl)-1-(2-amino-thiazol-4-yl)-3-(tetrahydro-4H-pyran-4-ylamino]-cyclopentanecarbamidehydrochloride salt (Example 197), 0.20 g of acetic anhydride and 0.40 gof pyridine in 1.0 mL of CH₂Cl₂ was stirred overnight, evaporated anddried in vacuo. The residue was purified by preparative TLC (10%[aqueous NH₄OH/MeOH 1/9]/CH₂Cl₂). The title compound (0.35 g, 59%) wasobtained as a mixture of cis and trans isomers. ¹H NMR (400 MHz, CDCl₃):1.50-3.40 (m, 20H), 4.00(m, 2H), 4.50 (m, 2H), 6.70, 7.05 (tt, 1H), 6.80(d, 1H), 7.60 (d, 2H), 7.75 (s, 1H), 9.10 (broad,1H). LC MS forC26H30F6N4O3S for [M+H]⁺ calc. 593, found 593.

EXAMPLE 199

Step A

A mixture of 0.65 g (2.0 mmol) of1-(2-tert-butoxycarbonyl-amino-thiazol-4-yl)-3-oxo-cyclopentanecarboxylicacid (Step G, Example 195), 0.65 g (2.5 mmol) of3-fluoro-5-trifluoromethylbenzylamine hydrochloride and 0.95 g EDC (5.0mmol) in 50 mL of CH₂Cl₂ was stirred for 2 h. The reaction mixture wasdiluted with 100 mL of CH₂Cl₂ and washed with 3 N aqueous HCl (3×50 mL),saturated aqueous NaHCO₃ (50 mL), water (100 mL), dried over Na₂SO₄ andevaporated in vacuo. 0.9 g of the title compound was obtained as ayellow solid. ¹H NMR (400 MHz, CDCl₃): 1.56 (s, 9H), 2.18 (m, 1H),2.38-2.65 (m, 3H), 2.70(d, 1H), 3.12 (d, 1H), 4.48 (m, 2H), 6.74 (s,1H), 7.10 (d, 1H—), 7.20-7.35 (m, 3H), 7.99 (broad, 1H). LC MS forC22H-23F4N₃O₄S for [M+H]⁺ calc. 502, found 502.Step B

This compound was prepared starting from the keto-amide (Step A, Example199) and Intermediate 2 according to the same procedure described underExample 195. LC MS for C27H34F4N4O4S for [M+H]⁺ calc. 587, found 587.

EXAMPLE 200

This compound was prepared starting from Example 199 according to thesame procedure described under Example 196. LC MS for C28H36F4N4O4S for[M+H]⁺ calc. 601, found 601.

EXAMPLE 201

This compound was prepared starting from Example 200 according to thesame procedure described under Example 197. LC MS for C23H28F4N4O2S for[M+H]⁺ calc. 501, found 501.

EXAMPLE 202

This compound was prepared starting from Example 201 according to thesame procedure described under Example 198. LC MS for C25H30F4N4O3S for[M+H]⁺ calc. 543, found 543.

EXAMPLE 203

Step A

A mixture of 1.1 g (2.0 mmol) ofN-(3,5-Bis-trifluoromethyl-benzyl)-1-(2-tert-butoxycarbonyl-amino-thiazol-4-yl)-3-oxo-cyclopentanecarbamide(Step H, Example 195) and 5 mL of neat TFA was stirred at roomtemperature for 1 h and evaporated. The residue was dissolved in 50 mLof EtOAc, washed with saturated aqueous sodium bicarbonate, dried overNa₂SO₄, evaporated and dried under vacuum. The title compound (0.85 g,94%) was obtained as a yellow solid. ¹H NMR (400 MHz, CDCl₃): 2.20 (m,1H), 2.38 (m, 1H), 2.52 (m, 2H), 2.60(d, 1H), 3.18 (d, 1H), 4.58 (m,2H), 5.34 (broad, 2H), 6.31 (s, 1H), 7.65 (2, 2H), 7.75 (s, 1H), 7.80(broad, 1H). LC MS for C18H15F6N3O2S for [M+H]⁺ calc. 452, found 452.Step B

A mixture of 0.85 g (1.9 mmol) ofN-(3,5-bis-trifluoromethyl-benzyl)-1-(2-amino-thiazol-4-yl)-3-oxo-cyclopentanecarbamide(Step A, Example 203), 1.0 mL of acetic anhydride and 2.0 mL of pyridinein 20 mL of CH₂Cl₂ was stirred overnight, diluted with 50 mL of CH₂Cl₂,washed with water and 2 N aqueous HCl, dried over Na₂SO₄ and evaporated.The title compound (0.74 g) was obtained as a light yellow solid afterpurification on prep TLC (10% MeOH/CH₂Cl₂). LC MS for C20H17F6N3O3S for[M+H]⁺ calc. 494, found 494.Step C

A mixture of 0.50 g (1.0 mmol) of,N-(3,5-bis-trifluoromethyl-benzyl)-1-(2-acetyl-amino-thiazol-4-yl)-3-oxo-cyclopentanecarbamide(Step B, Example 203), 0.27 g (2.0 mmol) of tetrahydro-4H-pyranylammonium chloride (Intermediate 2), 0.26 g (2.0 mmol) of DIEA, 0.84 g(4.0 mmol) of sodium triacetoxyborohydride and 0.5 g of molecular sieves(4 Å) in 20 mL of CH₂Cl₂ was stirred overnight, quenched with 50 mL ofsaturated aqueous Na₂CO₃. The solid was removed by filtration andwashing with CH₂Cl₂. Organic phase was separated and dried with Na₂SO₄,evaporated. The residue was purified by preparative TLC (10% [aq.NH₄OH/MeOH 1/9]/CH₂Cl₂). The title compound (0.34 g, 58%) was obtainedas a mixture of cis and trans isomers. LC MS for C25H28F6N4O3S for[M+H]⁺ calc. 579, found 579.

EXAMPLE 204

This compound, as a mixture of cis and trans isomers, was preparedstarting from the intermediate prepared in Step B, Example 203 andcyclohexyl amine according to the same procedure described under Step C,Example 203. LC MS for C26H30F6N4O2S for [M+H]⁺ calc. 577, found 577.

EXAMPLE 205

This compound, as a mixture of cis and trans isomers, was preparedstarting from the intermediate prepared in Step B, Example 203 andcyclopentyl amine according to the same procedure described under StepC, Example 203. LC MS for C25H28F6N4O2S for [M+H]⁺ calc. 563, found 563.

EXAMPLE 206

Step A

A mixture of 5.0 g (32 mmol) 2-methylthiazol-4-yl acetic acid, 2.4 g (50mmol) ethanol and 7.3 g (38 mmol) EDC in 50 mL of CH₂Cl₂ was stirred atroom temperature overnight. The entire mixture was loaded on a silicagel column and eluted with 30% EtOAc/hexane. The title compound wasobtained as a colorless oil. (4.5 g, 76% yield). ¹H NMR (300 MHz,CDCl₃): 1.24 (t, 3H), 2.66 (s, 3H), 3.75 (s, 2H), 4.15 (q, 4H), 6.98 (s,1H).Step B

To a mixture of 4.5 g (0.024 mol) of ethyl(2-methyl-thiazol-4-yl)acetate (Step A, Example 206),cis-1,4-dichloro-2-butene (3.2 mL, 0.030 mol) in 100 mL of DME at roomtemperature was added in multiple portions solid NaH (60% oil, 2.5 g,0.062 mol). The resulting mixture was stirred for 2 days, poured into300 mL of ice-water and extracted with 500 mL of ether. The ether layerwas washed with water (3×100 mL), dried over Na₂SO₄ and evaporated.Flash chromatography (silica gel, 10% EtOAc/hexane) afforded the titlecompound as an oil (3.0 g, 52%). ¹H NMR (400 MHz, CDCl₃): 1.20 (t, 3H),2.66 (s, 3H), 2.90 (d, 2H), 3.27 (d, 2H), 4.15 (q, 2H), 5.69 (s, 2H),6.87 (1, 1H).Step C

To a solution of 2.4 g (0.01 mol) of ethyl1-(2-methyl-thiazol-4-yl)-3-cyclopentenecarboxylate (Step B, Example206) in 30 mL of anhydrous ethyl ether at −78° C. was added dropwise asolution of BH₃.DMS in THF (5.0 mL, 0.015 mmol). The cooling bath wasremoved and the mixture was stirred at room temperature for 3 h, dilutedwith 100 mL of CH₂Cl₂, and 8.5 g of PCC was added. The mixture wasstirred overnight. The entire mixture was dumped onto a silica gelcolumn and eluted with 10% EtOAc/hexane. and then 30% EtOAc/hexane.Partial starting material (1.2 g, less polar) was recovered and thetitle compound (0.65 g, more polar) was obtained as an oil. ¹H NMR (300MHz, CDCl₃): 1.21 (t, 3H), 2.34 (m,2H), 2.58 (m,1H), 2.66 (s, 3H), 2.82(d, 1H), 3.05 (d, 2H), 4.19 (q, 4H), 6.95 (s, 1H).Step D

A mixture of 0.51 g (2.0 mmol) of ethyl1-(2-methyl-thiazolyl)-3-oxo-cyclopentanecarboxylate (Step C, Example206) and 200 mg (5 mmol) of lithium hydroxide monohydrate in a solutionof 5 mL of MeOH and 1 mL of water was stirred at room temperatureovernight, acidified with 2 N aqueous HCl, diluted with 50 mL of waterand extracted with ethyl acetate (2×50 mL). The combined organic layerswere dried over Na₂SO₄, filtered, evaporated and dried in vacuum. 0.12 gof the title product was obtained as a fluffy solid. ¹H NMR (300 MHz,CDCl₃): 2.30 (m, 1H), 2.22 (m, 1H), 2.50 (m, 1H), 2.65 (m, 1H), 2.72 (s,3H), 2.74 (d, 1H), 3.13 (d, 1H), 7.00 (s, 1H).Step E

A mixture of 0.12 g (0.50 mmol) of1-(2-methyl-thiazol-4-yl)-3-oxo-cyclopentanecarboxylic acid (Step D,Example 206), 0.2 g (0.7 mmol) of (3,5-bis-trifluoromethyl)benzylaminehydrochloride and 0.19 g EDC (1.0 mmol) in 5 mL of CH₂Cl₂ was stirredovernight. The reaction mixture was loaded on prep TLC and developedwith 10% MeOH/CH₂Cl₂. 0.17 g of the title compound was obtained as ayellow solid. ¹H NMR (400 MHz, CDCl₃): 2.18 (m, 1H), 2.40 (m, 1H), 2.60(m, 2H), 2.70 (s,3H), 2.77 (d, 1H), 3.25 (d, 1H), 4.58 (m, 2H), 6.97 (s,1H), 7.60 (s, 2H), 7.75 (s, 1H), 7.87 (broad, 1H). LC MS forC19H16F6N2O2S for [M+H]⁺ calc. 451, found 451.Step F

A mixture of 0.17 g (0.37 mmol) of,N-(3,5-bis-trifluoromethyl-benzyl)-1-(2-methyl-thiazol-4-yl)-3-oxo-cyclopentanecarbamide(Step E, Example 206), 0.27 g (2.0 mmol) of tetrahydro-4H-pyran-4-ylammonium chloride, 0.13 g (1.0 mmol) of DIEA, 0.8 g (4 mmol) of sodiumtriacetoxyborohydride and 0.5 g of molecular sieves (4 Å) in 30 mL ofCH₂Cl₂ was stirred overnight, quenched with 30 mL of saturated aqueousNa₂CO₃. The solid was removed by filtration and washing with CH₂Cl₂. Theorganic phase was separated and dried with Na₂SO₄ and evaporated. Theresidue was purified by preparative TLC (10% [aqueous NH₄OH/MeOH1/9]/CH₂Cl₂). The title compound (0.11 g, 56%) was obtained as a mixtureof cis and trans isomers. Further separation on prep LC (10%MeOH/CH₂Cl₂) afforded two components. Less polar (trans isomer) ¹H NMR(400 MHz, CDCl₃): 1.30-1.35 (m, 3H), 1.80 (m, 3H), 2.10 (m, 1H), 2.30(m, 1H), 2.50 (m, 1H), 2.67 (s, 3H), 2.75 (m, 1H), 2.92 (m, 1H), 3.40(t, 3H), 3.58 (m, 1H), 3.98 (m, 2H), 4.56 (m, 2H), 7.00 (s, 1H), 7.46(broad, 1H), 7.54 (s, 2H), 7.73 (s, 1H). LC MS for C24H27F6N3O2S for[M+H]⁺ calc. 563, found 563.

More polar (cis isomer) ¹H NMR (400 MHz, CDCl₃): 1.50 (m, 2H), 1.65 (m,1H), 1.80 (m, 2H), 2.10 (m, 1H), 2.20 (m, 1H), 2.50 (d, 2H), 2.60 (m,1H), 2.69 (s, 3H), 2.82 (m, 1H), 3.38 (t, 2H), 3.50 (m, 1H), 3.98 (m,2H), 4.58 (m, 2H), 6.99 (s, 1H), 7.57 (s, 2H), 7.76 (s, 1H), 7.84(broad, 1H). LC MS for C24H27F6N3O2S for [M+H]⁺ calc. 563, found 563.

EXAMPLE 207

This compound, as a mixture of cis and trans isomers, was preparedstarting from 2-thiazol-4-yl acetic acid following the same proceduredescribed under Example 206. LC MS for C23H25F6N3O2S for [M+H]⁺ calc.522, found 522.

EXAMPLE 208

Step A

To a stirred solution of phenyl magnesium bromide (3 M solution inether, 680 mL, 2.05 mol) in ethyl ether (500 mL) was addedexo-epoxynorbornane (150 g, 1.36 mol) in ethyl ether (250 mL) slowly.After the initial exotherm, the reaction was heated to reflux for 3 h,after which time it was cooled in an ice bath and quenched with water(25 mL). The resulting solution was diluted with ethyl ether and washedwith aqueous 3 N HCl twice. The combined aqueous layers where backextracted with ethyl ether twice and the combined organic layers wherewashed with brine, dried over MgSO₄, filtered, and concentrate underreduced pressure (100 mmHg, 30° C.) to give 230 g of a crude orange oil.This material was subject to flash chromatography (silica gel, 40% ethylether/hexanes) to give 67 g of pure product (45%). ¹H NMR (CDCl₃, 500MHz): 6.06 (d, J=1.0 Hz, 2H), 3.76 (s, 1H), 2.75 (d, J=2.0 Hz, 2H), 1.86(bs, 2H), 1.71-1.68 (m, 2H).Step B

To a cooled (−78° C.) solution of oxalyl chloride (83 g, 660 mmol) inDCM (500 mL) was added DMSO (78 mL, 1.1 mol) in DCM (200 mL) rapidly butkeeping the temperature below −50° C. To this solution was immediatelyadded the product from Step A (67 g, 610 mmol) in DCM (600 mL) rapidly,but keeping the temperature below −50° C. After stirring for 15 minutesat −78° C. this solution was treated with triethylamine (310 mL, 2.1mol) and allowed to warm to room temperature. After 1 h at roomtemperature, the reaction was quenched with water and concentrated underreduced pressure. The crude residue was dissolved in a 3:1 solution ofethyl ether and petroleum ether and washed 3 times with aqueous 1 N HClthen with brine. The organic layer was dried over Na₂SO₄, filtered, andconcentrated under reduced pressure. The resulting residue was quicklychromatographed (short column—silica gel, 15% ethyl ether/hexanes) andconcentrated under reduced pressure. Final purification was achieved bydistillation (collecting the 60° C. to 70° C. fractions at 30 mm Hg) togive 18.5 g of pure product as a colorless liquid (28%). ¹H NMR (CDCl₃,500 MHz): 6.53 (bs, 2H), 2.82 (bs, 2H), 1.97 (d, J=7.0 Hz, 2H), 1.21(dd, J=4.5, 6.5 Hz, 2H).Step C

The product from Step B (17.5 g, 162 mmol) was combined withp-toluenesulfonic acid (4.9 g, 26 mmol) and ethylene glycol (13.1 mL,243 mmol) in benzene (200 mL) and heated to reflux. After 5 h, thesolution was allowed to cool to room temperature and stir overnight,after which time it was partitioned between ethyl ether and aqueoussaturated NaHCO₃. The organic phase was washed with brine, dried overMgSO₄, filtered and concentrated. The product was purified by flashchromatography (silica gel, 10% ethyl ether/hexanes) to give 19.0 g of acolorless oil (83%). ¹H NMR (CDCl₃, 500 MHz): 6.18 (bs, 2H, 3.92 (t,J=6.0 Hz, 2H), 3.85 (t, J=6.0 Hz, 2H), 2.53 (bs, 2H), 1.92 (d, J=7.5 Hz,2H), 0.97 (dd, J=3.5, 10.5 Hz, 2H).Step D

A solution of the product from Step C (2.0 g, 13 mmol) in a mixture ofmethanol (30 mL) and DCM (24 mL) was cooled to −78° C. and treated withozone gas (7.5 psi, 2 L/min) until a blue tint to the solution wasapparent. At this time, the reaction was purged with nitrogen gas toremove the excess ozone and sodium borohydride (600 mg, 16 mmol) wasadded to the reaction. The reaction was allowed to warm to 0° C. on anice bath before acetone was added to quench the excess reducing agent.The resulting solution was concentrated under reduced pressure and theproduct was purified by flash chromatography (silica gel, eluting withEA) to give 1.9 g of a colorless oil which upon cooling to −20° C.became a colorless solid (78%). ¹H NMR (CDCl₃, 500 MHz): 4.02 (m, 4H),3.67 (m, 4H), 2.22 (t, J=6.0 Hz, 2H), 1.83 (m, 2H), 1.63 (m, 2H).Step E

To a cooled (−15° C.) solution of the product from Step D (1.26 g, 6.71mmol) in THF (21 mL) was added n-butyllithium (2.5 M in hexanes, 2.8 mL,7.0 mmol). After the reaction was stirred for 30 minutes at −15° C.,tosyl chloride (1.28 g, 6.71 mmol) in THF (10 mL) was added dropwise andthe reaction was warmed to room temperature and stirred for anadditional 30 minutes before being concentrated under reduced pressure.The mono-tosylate product was separated from small amounts of startingmaterial and di-tosylation product by medium pressure liquidchromatography (silica gel, 40-100% EA/hexanes) to give 900 mg of acolorless oil (39%) which was used directly in the next step.Step F

The product from Step E (707 mg, 2.07 mmol) was combined with sodiumhydride (60% dispersion in mineral oil, 250 mg) in THF and stirred atroom temperature. After 2 h the reaction was quenched with hydrogenchloride (2 N solution in ethyl ether, 4 mL) and the resultingprecipitate was filtered off. The filtrate was concentrated and purifiedby flash chromatography (silica gel, 20% ethyl ether/hexanes) to give320 mg of product (91%). ¹H NMR (CDCl₃, 500 MHz): 3.97 (m, 4H), 3.93 (d,J=10.5 Hz, 2H), 3.57 (dd, J=2.5, 11.0 Hz, 2H), 1.84-1.81 (m, 2H), 1.75(m, 4H).Step G

The product from Step F (250 mg, 1.47 mmol) was dissolved in a mixtureof THF (4 mL) and aqueous 5% HCl (2 mL) and stirred at room temperature.After 18 h the reaction was diluted with ethyl ether, washed with brine,and dried over MgSO₄, filtered and concentrated under reduced pressure.The product was purified by flash chromatography (silica gel, 30% ethylether/hexanes) to give 51 mg of a volatile liquid (28%). ¹H NMR (CDCl₃,500 MHz): 3.99 (dd, J=2.5, 11.0 Hz, 2H), 3.87 (d, J=1 Hz, 2H), 2.28 (bs,2H), 2.03 (m, 2H), 1.99 (m, 2H).Step H

Intermediate 16 (30 mg, 0.07 mmol) was combined with the product fromStep G (9 mg, 0.07 mmol), N,N-diisopropylethylamine (25 μL, 0.15 mmol),4 Å powdered molecular sieves (50 mg), and sodium triacetoxyborohydride(75 mg, 0.35 mmol) in 5 mL DCM. The reaction mixture was stirred at roomtemperature for 6 days, then filtered through celite, and concentratedunder reduced pressure. The product was purified by preparative TLC(silica gel, 0.3% NH₄OH/3.7% MeOH (97% DCM) and converted to itshydrochloride salt by the addition of hydrogen chloride (2 N solution inethyl ether) to give 9 mg of a white solid (25%). ESI-MS calc. forC25H32F6N2O2: 506; found 507 (M+H).

EXAMPLE 209

Step A:

Intermediate 19 (860 mg, 4.8 mmol) was added to a stirred solution of1,2,4-triazole (280 mg, 4.0 mmol) in THF (5 mL) and DMF (5 mL). K₂CO₃(690 mg, 5.0 mmol) was added to the solution and the reaction wasallowed to stir at room temperature for 18 h at which time it wasfiltered and the concentrated to dryness. The product was purified bymedium pressure liquid chromatography (silica gel, 10-100% EA/hexanes)to give 430 mg of a single isomer (54%). ¹H NMR (CDCl₃, 500 MHz) 8.26(s, 1H), 7.98 (s, 1H), 5.18 (dd, J=6.5, 17.5 Hz, 1H), 4.57 (m, 1H), 4.37(m, 1H), 4.05 (t, J=10.5 Hz, 1H), 3.86 (m, 1H), 2.84 (m, 1H), 2.69 (m,1H).Step B

The product from Step A (24 mg, 0.15 mmol) was added to a solution ofIntermediate 16 (52 mg, 0.12 mmol) and triethylamine (17 μL, 0.12 mmol)in DCM (5 mL). After 5 minutes at room temperature this solution wastreated portionwise with 4 Å molecular sieves (100 mg) and sodiumtriacetoxyborohydride (100 mg). After 72 h at room temperature thereaction was treated with an additional amount of the product from StepA (24 mg, 0.15 mmol) and allowed to stir for an additional 72 h. Thereaction was filtered through celite washing with DCM, and concentratedunder reduced pressure. The product was purified by preparative TLC(silica gel, 1000 gm, 0.5% NH₄OH/4.5% MeOH/95% DCM) and converted to itshydrochloride salt by the addition of 2 N HCl in ethyl ether to give 22mg of a white solid (30%). ESI-MS calc. for C25H31F6N5O2: 547; found 548(M+H).

EXAMPLE 210

Step A

Intermediate 19 (860 mg, 4.8 mmol) was added to a stirred solutiontetrazole (280 mg, 4.0 mmol) in THF (5 mL) and DMF (5 mL). K₂CO₃ (690mg, 5.0 mmol) was added to the solution and the reaction was allowed tostir at room temperature for 18 h at which time it was filtered and theconcentrated to dryness. The product was purified by medium pressureliquid chromatography (silica gel, 30-100% EA/hexanes) to give 260 mg ofa single isomer (38 to). ¹H NMR (CDCl₃, 500 MHz) 8.85 (s, 1H), 5.51 (dd,J=6.5, 17.5 Hz, 1H), 4.75 (m, 1H), 4.45 (m, 1H), 4.00 (t, J=9.5 Hz, 1H),3.86 (m, 1H), 2.93 (m, 1H), 2.75 (m, 1H).Step B

The product from Step A (25 mg, 0.15 mmol) was added to a solution ofIntermediate 16 (52 mg, 0.12 mmol) and triethylamine (17 μL, 0.12 mmol)in DCM (5 mL). After 5 minutes at room temperature this solution wastreated portionwise with 4 Å molecular sieves (100 mg) and sodiumtriacetoxyborohydride (100 mg). After 72 h at room temperature thereaction was treated with an additional amount of the product from StepA (24 mg, 0.15 mmol) and allowed to stir for an additional 72 h. Thereaction was filtered through celite, washing with DCM, and concentratedunder reduced pressure. The product was purified by preparative TLC(silica gel, 1000 μm, 0.5% NH₄OH/4.5% methanol/95% DCM) and converted toits hydrochloride salt by the addition of 2 N HCl in ethyl ether to give1.8 mg of a white solid (2%). ESI-MS calc. for C24H30F6N6O2: 548; found549 (M+H).

EXAMPLE 211

Step A

A cooled (−78° C.) THF (80 mL) solution of commercially availablemethyl-(3-methylenecyclopentane) carboxylate (3.90 g, 27.8 mmol) wastreated dropwise with 1.5 M LDA in cyclohexane (22.3 mL, 33 mmol) over10 minutes. The reaction mixture was stirred for an additional 35minutes., then a solution of 4-bromocrotononitrile (1:2 trans/cis,prepared according to Zindel, J.; de Meijere, A., Synthesis (1994),190-194. 4.26 g, 29.2 mmol) in THF (5 mL) was added dropwise over 10minutes. The reaction mixture was stirred at −78° C. for 1.5 h., thenpoured into a 10% aqueous citric acid solution. This mixture wasextracted twice with ether (300 mL), the ethereal layers were combined,and these in turn were washed with saturated NaHCO₃ solution, followedby brine. The ethereal layer was then dried over anhydrous MgSO₄,filtered, and concentrated. Purification by MPLC (silica, 40%ether/hexane) afforded two product mixtures (6:1 ratio), correspondingto the four isomers with trams-cyclopropyl stereochemistry (3.07 g) andthe four with cis-cyclopropyl stereochemistry (504 mg), respectively.Step B

Ozone gas was bubbled through a cooled solution (−78° C.) of the olefinprepared as described in Step A above (top spot, trans-cyclopropyl, 3.07g, 15.0 mmol) in DCM (50 mL) until the reaction mixture became blue incolor. Then nitrogen gas was bubbled through the solution until it wascolorless again. Triphenylphosphine (4.33 g, 16.5 mmol) was then addedand the reaction mixture was permitted to warm to room temperature andstir for three h. The reaction mixture was then concentrated andpurified by flash chromatography (silica, eluted with DCM, then 1%methanol/DCM, then 3% methanol/DCM) to give 1.31 g of product as amixture of 4 diastereomers (trans cyclopropyl). The olefin prepared inStep A having the cis-cyclopropyl stereochemistry (bottom spot) wasconverted to its corresponding ketone in the same fashion as thatdescribed immediately above.Step C

The trans-cyclopropyl ketone from Step B above (790 mg, 3.8 mmol) wasdissolved in 1:1 THF/methanol (16 mL) and treated with a solution ofLiOH.H₂O (800 mg, 19 mmol) in 8 mL of water. The resulting reactionmixture was stirred at room temperature for 1 h, then neutralized with 3N HCl solution, and concentrated to remove the organic solvents. Theaqueous product mixture was then extracted three times with chloroform,the organic layers were combined and dried over anhydrous MgSO₄,filtered, and concentrated to give 604 mg of carboxylic acid product(82%).Step D

The trans-cyclopropyl keto-acid from Step C above (570 mg, 2.9 mmol) wasadded to a stirred solution of1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (1.1 g, 5.9 mmol) in DCM(30 mL). To this was added 3,5-bistrifluorobenzylamine hydrochloride(1.2 g, 4.4 mmol), triethylamine (440 mg, 4.4 mmol), andN,N-dimethylamino pyridine (5 mg). The reaction was stirred at roomtemperature for 18 h before being diluted with DCM and washed threetimes with aqueous 1 N HCl, once with water, three times with saturatedaqueous sodium bicarbonate, and once with brine. The organic layer wasdried over MgSO₄, filtered and concentrated under reduced pressure. Theproduct was purified by medium pressure liquid chromatography (silicagel, 70% EA/Hexanes) to give 229 mg of a colorless oil. ESI-MS calc. forC19H16F6N2O2: 418; found 419 (M+H).Step E

The trans-cyclopropyl keto-amide from Step D above (200 mg, 0.47 mmol)was combined with 4-amino-4H-tetrahydropyran (78 mg, 0.57 mmol),triethylamine (57 mg, 0.56 mmol), 4° A powdered molecular sieves (100mg), and sodium triacetoxyborohydride (400 mg, 1.9 mmol) in 5 mL DCM.The reaction mixture was stirred at room temperature for 3 days, thenfiltered through celite, diluted with DCM, and washed with saturatedNaHCO₃ solution and brine. The organic layer was dried over anhydrousMgSO₄, filtered and concentrated. The product was purified by reversephase HPLC (C18, 20-100% MeCN/H₂O) to give 119 mg of a mixture of 8isomers. Preparatory TLC (0.5% NH₄OH/4.5% MeOH/95% DCM) of this productgave 2 separate mixtures of 4 isomers. 40 mg of the top spot,cis-racemate cyclopentyl/trans-cyclopropyl was recovered: ESI-MS calcfor C24H27F6N3O2: 503; found 504 (M+H). 5.6 mg of the bottom spot,trans-racemate cyclopentyl/trans-cyclopropyl was recovered: ESI-MS calcfor C24H27F6N3O2: 503; found 504 (M+H).

EXAMPLE 212

N-methylation of Example 211 (34 mg, 0.068 mmol) was carried outaccording to the standard procedure (Example 2) to give 21 mg of product(61%). ESI-MS calc. for C25H29F6N3O2: 517; found 517 (M+H).

EXAMPLE 213

Step A

Intermediate 37 (210 mg, 1.7 mmol) was dissolved in THF (10 mL) andcooled to −78° C. A 1.0 M solution of LS-Selectride in THF (3.4 mL, 3.4mmol) was added dropwise and the solution was allowed to warm to roomtemperature over 4 h at which time it was left at room temperature for12 h before being concentrated under reduced pressure. The resulting oilwas dissolved in DCM, washed with an aqueous 1 N HCl solution, then withbrine, dried over MgSO₄, filtered and concentrated. The product waspurified by flash chromatography (silica, 45% petroleum ether/ethylether) to give 144 mg of a colorless oil (66%). ¹H NMR (CDCl₃, 500 MHz):4.35 (bs, 2H), 4.12 (t, J=4.5 Hz, 1H), 2.24-2.13 (m, 2H), 2.12-2.08 (m,2H), 1.98-1.90 (m, 2H), 1.63 (d, J=11.5 Hz, 2H).Step B

To a cooled (0° C.) solution of the product from Step A (120 mg, 0.95mmol), triethylamine (144 μL, 1.04 mmol) and DMAP (2-3 crystals) in DCM(10 mL), was added methane sulfonyl chloride (80 μL, 1.0 mmol) dropwise.The solution was stirred at 0° C. for 2.5 h before an additional portionof methane sulfonyl chloride (80 μL, 1.0 mmol) was added. The solutionwas warmed to room temperature and stirred for 16 h. The reaction waswashed with saturated sodium bicarbonate solution, washed with brine,dried over MgSO₄, filtered and concentrated to give 205 mg of a yellowoil (99%+). The product was used in the next step without furtherpurification.Step C

To a solution of the product form Step B (112 mg, 0.546 mmol) in DMSO (2mL) was added sodium azide (177 mg, 2.73 mmol). The solution was heatedto 50° C. for 18 h then diluted with DCM, washed with water twice,washed with brine, dried over MgSO₄, filtered and concentrated to give80 mg of a yellow oil (96%). ¹H NMR (CDCl₃, 500 MHz): 4.48 (bs, 2H),3.66-3.60 (m, 1H), 2.64 (bs, 2H), 2.04-1.98 (m, 2H), 1.85-1.77 (m, 2H),1.75-1.65 (m, 2H).Step D

To a solution of the product from Step C (80 mg, 0.54 mmol) in methanol(2 mL) was added 20% Pd(OH)₂ (w/w) on C (16 mg). The solution was placedunder a hydrogen balloon for 5.5 h. The solution was filtered throughcelite and concentrated to give 44 mg of a yellow oil (65%).Step E

The product from step D (44 mg, 0.35 mmol) was combined withIntermediate 8 (100 mg, 0.25 mmol), 4 Å powdered molecular sieves (˜200mg), and sodium triacetoxyborohydride (212 mg, 1.00 mmol) in DCM (5 mL).The resulting mixture was stirred at room temperature for 5 d. Thereaction mixture was filtered through celite and concentrated. Theproduct was purified by preparative TLC (silica, 7% NH₃/MeOH in DCM) togive a top and bottom spot. The bottom spot was converted to its HClsalt by addition of a solution of HCl in dioxane to give 31.2 mg of awhite solid (25%). The top spot was further purified by preparative TLC(silica, 7% NH₃/MeOH in DCM) and converted to its HCl salt by theaddition of a solution of HCl in dioxane to give 28.6 mg of a whitesolid (20%).

Top Spot, cis racemate: ESI-MS calc. for C25H32F6N2O2: 506; Found: 507(M+H).

Bottom Spot, trans racemate: ESI-MS calc. for C25H32F6N2O2: 506; Found:507 (M+H).

EXAMPLE 214

Intermediate 37 (43 mg, 0.35 mmol) was combined with Intermediate 16(100 mg, 0.23 mmol), triethylamine (32 μL, 0.23 mmol), 4 Å powderedmolecular sieves (50 mg), and sodium triacetoxyborohydride (200 mg, 0.92mmol) in 10 mL of DCM. The reaction mixture was stirred at roomtemperature for 3 days, then filtered through celite, diluted with DCM,and washed with a saturated NaHCO₃ solution and brine. The organic layerwas dried over anhydrous MgSO₄, filtered and concentrated. The productwas purified preparative TLC (0.7% NH₄OH (6.3% MeOH/93% DCM) andconverted to its hydrochloride salt by addition of 2 N HCl in ethyl togive 47 mg of a white solid as an unknown endo/exo mixture. ESI-MS calc.for C25H32F6N2O2: 506; Found: 507 (M+H).

EXAMPLE 215

Step A

To a cooled (−78° C.) solution of LDA (1.5 M solution in cyclohexane, 16mL, 24 mmol) in THF (80 mL) was added a solution oftetrahydro-4H-pyran-4-one (2 g, 20 mmol) and HMPA (3.6 mL, 21 mmol) inTHF (20 mL). After 1 h at −78° C. the solution was treated with benzylbromide (4.7 mL, 40 mmol) and allowed to warm to room temperature whereit was allowed to stir overnight. The reaction was poured over ice andextracted 3 times with diethyl ether. The combined organic layers wherewashed with brine, dried over MgSO₄, filtered, and concentrated underreduced pressure. The product was purified by flash chromatography(silica gel, 30% ether/hexanes) to give 800 mg of the desiredmono-alkylation product and 1.16 g of an un-desired di-alkylationproduct. ¹H NMR (desired mono-alkylation product): (CDCl₃, 500 MHz)7.35-7.15 (m, 5H), 4.21 (m, 1H), 4.06 (dd, J=5.5, 11.5 Hz, 1H), 3.81 (t,J=10 Hz), 3.22 (dd, J=5.0, 15.5 Hz, 1H), 2.85 (m, 1H), 2.63 (m, 1H),2.50 (m, 2H).Step B

The product from Step A (88 mg, 0.46 mmol) was combined withIntermediate 16 (100 mg, 0.23 mmol), triethylamine (32 μL, 0.23 mmol), 4Å powdered molecular sieves (60 mg), and sodium triacetoxyborohydride(240 mg, 1.1 mmol) in 10 mL DCM. The reaction mixture was stirred atroom temperature for 3 days, then filtered through celite, diluted withDCM, and washed with saturated NaHCO₃ solution and brine. The organiclayer was dried over anhydrous MgSO₄, filtered and concentrated. Theproduct was purified preparative TLC (0.7% NH₄OH/6.3% MeOH/93% DCM) andconverted to its hydrochloride salt by the addition of 2 N HCl in ethylether to give 6 mg of a white solid as a mixture of 4 isomers. ESI-MScalc. for C30H36F6N2O2: 570; Found: 571 (M+H).

EXAMPLE 216

Example 128 was resolved into its 4 single diastereomers using aChiralCel OD column, eluting with 3% ethanol/hexanes.

241A: First peak: ESI-MS calc. for C24H32F6N2O2: 494; found 495 (M+H).

241B: Second peak: ESI-MS calc. for C24H32F6N2O2: 494; found 495 (M+H).

241C: Third peak: ESI-MS calc. for C24H32F6N2O2: 494; found 495 (M+H).

241D: Fourth peak: ESI-MS calc. for C24H32F6N2O2: 494; found 495 (M+H).

EXAMPLE 217

The Top Spot from Example 154 was resolved into 3 single diastereomersusing a ChiralCel OD column, eluting with 20% isopropanol/hexanes.

242A: ESI-MS calc. for C26H34F6N2O2: 520; found 521 (M+H).

242B: ESI-MS calc. for C26H34F6N2O2: 520; found 521 (M+H).

242C: ESI-MS calc. for C26H34F6N2O2: 520; found 521 (M+H).

EXAMPLE 218

N-methylation of Example 242C (5.0 mg, 0.01 mmol) was carried outaccording to the standard procedure (see Example 2) to give 1.4 mg ofproduct (28%). ESI-MS calc. for C27H36F6N2O2: 534; found 535 (M+H).

EXAMPLE 219

Step A

A mixture of 3-bromo-5-trifluoromethylbenzonitrile (9.9 g, 41 mmol),zinc cyanide (9.7 g, 82 mmol), and Pd(PPh₃)₄ (2.4 g, 2 mmol) inanhydrous N,N-dimethylformamide (120 ml) was degassed then heated at 90°C. for 6 h. The cooled reaction mixture was poured into water (600 ml)and extracted with ethyl acetate (3×150 ml). The combined ethyl acetatelayers were washed with water (3×200 ml), saturated NaCl (100 ml), driedover MgSO₄, filtered and evaporated. The residue was purified by columnchromatography on silica eluting with 20% ethyl acetate in hexanes togive the desired product (6.31 g, 84%); ¹H NMR 500 MHz (CDCl₃): 4.21(2H, s), 7.05 (1H, s), 7.09 (1H, s), 7.23 (1H, s).Step B

A solution of the product from Step A (4.75 g, 25.5 mmol) in a mixtureof ethanol (50 mL) and ammonium hydroxide (10 mL) was hydrogenated at 40psi over Raney Nickel (0.5 g) for 18 h on a Parr apparatus. The catalystwas removed by filtration thru celite, and the filtrate was evaporatedto dryness. The resultant residue was dissolved in DCM, dried overMgSO₄, filtered and treated with di-tert-butyl dicarbonate (6.12 g, 28.0mmol). The reaction was stirred at room temperature for 3 h beforeN,N-dimethyl ethylenediamine was added. After 30 minutes, the reactionwas washed with aqueous 1 M citric acid, followed by aqueous saturatedNaHCO₃, and brine. The organic layer was dried over MgSO₄, filtered andconcentrated to give 4.5 g of product (62%). ¹H NMR (CDCl₃, 500 MHz):6.85 (s, 1H), 6.75 (s, 1H), 6.70 (s, 1H), 5.10 (bs, 1H), 4.20 (bs, 2H),3.85 (bs, 2H), 1.60-1.20 (m, 9H).Step C

To a mixture of the product from Step B (3.5 g, 12 mmol) andN,N-dimethyl formamide azine (3.5 g) in toluene (100 mL) was addedp-toluene sulfonic acid, and the resulting mixture was heated to reflux.After 18 h at reflux the reaction was cooled, washed with saturatedaqueous NaHCO₃, dried over K₂CO₃, filtered, and concentrated underreduced pressure. The residue was purified using a Biotage flash 40(silica gel, 0.5% NH₄OH/4.5% methanol/DCM) to give 2.1 g of product(51%). ESI-MS calc. for C15H17F3N4O2: 342.1; found 343.1 (M+H).Step D

To a solution of the product from Step C (2.1 g, 6.1 mmol) in DCM (20mL) was added trifluoroacetic acid (5 mL) and the mixture was stirred atroom temperature. After 2 h the reaction was concentrated under reducedpressure and the residue was made alkaline with saturated aqueous NaHCO₃and washed three times with DCM. The aqueous layer was evaporated todryness and the resulting residue was triturated with methanol. Themethanol triturate was evaporated under reduced pressure, and the crudeproduct was purified by reverse phase HPLC (C18, 20-100% MeCN/H₂O). Theresulting TFA salt was loaded onto Dowex 50 W×8 resin and washed withwater then eluted with 10% aqueous NH₄OH to give 700 mg of a yellow oil(47%). ¹H NMR (CDCl₃, 500 MHz): 9.22 (s, 2H), 8.60 (bs, 2H), 8.20 (s,1H), 8.19 (s, 1H), 7.97 (s, 1H), 4.20 (s, 2H).Step E

Intermediate 17 (700 mg, 2.6 mmol) was dissolved in DCM (10 mL) andtreated with oxalyl chloride (230 μL, 5.2 mmol) and 1 drop of DMF. After2 h at room temperature the reaction was concentrated to dryness anddried for 1.5 under high vacuum. 160 mg of this acid chloride (0.56mmol) was dissolved in DCM (5 mL) and added dropwise to a stirredsolution of the product from Step D (70 mg, 0.29 mmol) in triethylamine(2 mL). After 15 minutes an additional 100 mg of the acid chloride wasadded to the reaction and stirring was continued at room temperature.After 1 h the reaction was concentrated under reduced pressure andpassed through a Spe-ed SCX column, washing with methanol and elutingwith 2 M NH₃ solution in methanol. This crude product was twice furtherpurified by reverse phase HPLC (C18, 25-100% MeCN/H₂O) and converted toits hydrochloride salt by the addition of hydrogen chloride (2 Nsolution in ethyl ether) to give 13 mg of a white solid (9%). ESI-MScalc. for C25H34F3N5O2: 493; found 494 (M+H).

EXAMPLE 220

Step A

A solution of the iodo-nitrile prepared in Step A of Intermediate 7 (220mg, 0.74 mmol) in toluene (2.5 mL) was treated with sodium azide (160mg, 2.5 mmol) and triethylamine hydrochloride (350 mg, 2.5 mmol) andheated to 100° C. After 18 h the reaction was cooled to room temperatureand extracted twice with H₂O. The aqueous layer was acidified withconcentrated aqueous HCl, and the resulting precipitate was filtered,washed with water and dried under high vacuum to give 250 mg of a whitesolid (99%). ESI-MS calc. for C8H4F31N4: 340; found 341.Step B

The product from Step A (230 mg, 0.68 mmol) was combined withtetrakis(triphenylphosphine) palladium (47 mg, 0.041 mmol) and zinccyanide (110 mg, 0.95 mmol) in DMF (deoxygenated) and heated at 80°.After 18 h, an additional 50 mg of zinc cyanide was added and thereaction was stirred at 80° C. for an additional 3 h before being cooledto room temperature and extracted twice with aqueous 2 N NH₄OH. Thecombined aqueous layers were acidified with concentrated aqueous HCl andextracted 4 times with ethyl ether. The combined organic layers wherewashed with brine, dried over MgSO₄, filtered and concentrated underreduced pressure. The product was purified by medium pressure liquidchromatography (silica gel, 50 to 100% EA/hexanes) to give 100 mg of anoil (62%). ESI-MS calc. for C9H4F3N5: 239; found 240.Step C

The product from Step B (90 mg, 0.38 mmol) was dissolved in THF (3 mL)and treated with borane (1.0 M solution in THF, 3.8 mL, 3.8 mmol). After18 h at room temperature, the reaction was quenched with a 1% hydrogenchloride solution in methanol (10 mL) and heated to 50° C. After 18 h.the reaction was concentrated under reduced pressure and the residue wasre-dissolved in a 1% hydrogen chloride solution in methanol (10 mL).After 4 h the solution was concentrated under reduced pressure to give120 mg of product which was used directly in the next step. ESI-MS calc.for C9H8F3N5: 243; found 244 (M+H)Step D

Intermediate 17 (700 mg, 2.6 mmol) was dissolved in DCM (10 mL) andtreated with oxalyl chloride (230 μL, 5.2 mmol) and 1 drop of DMF. After2 h at room temperature the reaction was concentrated to dryness anddried for 1.5 under high vacuum. 20 mg of this acid chloride (0.07 mmol)was dissolved in DCM (1 mL) and added dropwise to a stirred solution ofthe product from Step C (10 mg, 0.04 mmol) in triethylamine (1 mL).After 18 h the reaction was concentrated under reduced pressure andpassed through a Spe-ed SCX column, washing with methanol and elutingwith 2 M NH₃ solution in methanol. This crude product was furtherpurified by reverse phase HPLC (C18, 25-100% MeCN/H₂O) and converted toits hydrochloride salt by addition of hydrogen chloride (2 N solution inethyl ether) to give 10 mg of a white solid (50%). ESI-MS calc. forC24H33F3N6O2: 494; found 495 (M+H).

EXAMPLE 221

Step A

To a cooled (0° C.) solution of the iodo-nitrile prepared in Step A ofIntermediate 7 (500 mg, 1.7 mmol) in DMSO (2 mL) was added with asolution of K₂CO₃.1.5H₂O (33 mg, 0.20 mmol) and H₂O₂ (30% solution inwater, 340 μL, 3.1 mmol) in water (4 mL) and the resulting solution wasallowed to warm to room temperature. After 18 h additional quantities ofboth K₂CO₃ (280 mg, 1.7 mmol) and H₂O₂ (610 μL, 5.4 mmol) where addedand the reaction was stirred at room temperature for an additional 30minutes. The reaction was diluted with water (10 mL) and extracted withEA. The organic layer was washed twice with water, then brine, and driedover MgSO₄, filtered and concentrated under reduced pressure to give 450mg of product (85%).Step B

The product from Step A (410 mg, 1.3 mmol) was dissolved inN,N-dimethylformamide dimethyl acetal (10 mL) and heated to 120° C. for4 h before the N,N-dimethylformamide dimethyl acetal was distilled offunder reduced pressure. The resulting residue was treated with asolution of hydrazine hydrate (76 μL, 1.6 mmol) in glacial acetic acid(10 mL) and heated to 90° C. After 1 h the reaction was concentratedunder reduced pressure to give 442 mg of product (99%). ESI-MS calc. forC9H5F31N3: 339; found 340 (M+H).Step C

The product from Step B (440 mg, 1.3 mmol) was combined withTetrakis(triphenylphosphine) palladium (90 mg, 0.078 mmol) and zinccyanide (300 mg, 2.6 mmol) in DMF (deoxygenated) and heated to reflux.After 110 h, the reaction was cooled to room temperature and partitionedbetween ethyl ether and aqueous 2 N NH₄OH. The aqueous layer wasextracted twice with ether and the combined organic layers where washedwith brine, dried over MgSO₄, filtered and concentrated under reducedpressure to give 350 mg of product which was used directly in the nextstep. ESI-MS calc. for C10H5F3N4: 238; found: 239 (M+H).Step D

The product from Step C (350 mg, 1.5 mmol) was dissolved in THF (10 mL)and treated with borane (1.0 M solution in THE, 7.4 mL, 7.4 mmol). After2 h at room temperature, the reaction was quenched with a 1% hydrogenchloride solution in methanol (20 mL) and heated to 50° C. After 18 h.the reaction was concentrated under reduced pressure and the residue wasre-dissolved in a 1% hydrogen chloride solution in methanol (20 mL).After 2 h the solution was concentrated under reduced pressure to give410 mg a white salt (98%). ESI-MS calc. for C10H9F3N4: 242; found 243(M+H).Step E

Intermediate 17 (700 mg, 2.6 mmol) was dissolved in DCM (10 mL) andtreated with oxalyl chloride (230 μL, 5.2 mmol) and 1 drop of DMF. After2 h at room temperature the reaction was concentrated to dryness anddried for 1.5 under high vacuum. 90 mg of this acid chloride (0.31 mmol)was dissolved in DCM (2 mL) and added dropwise to a stirred solution ofthe product from Step D (45 mg, 0.19 mmol) in triethylamine (2 mL).After 3.5 h the reaction was concentrated under reduced pressure andpassed through a Spe-ed SCX column, washing with methanol and elutingwith 2 M NH₃ solution in methanol. This crude product was furtherpurified by reverse phase HPLC (C18, 25-100% MeCN/H₂O) and converted toits hydrochloride salt by the addition of hydrogen chloride (2 Nsolution in ethyl ether) to give 3.7 mg of a white solid (4%). ESI-MScalc. for C25H34F3N5O2: 493; found 494 (M+H).

EXAMPLE 222

A mixture of the Intermediate 8 (79 mg, 0.20 mmol), cyclohexylamine (46μL, 0.40 mmol), molecular sieve (4 Å, 90 mg), DIEA (70 μL, 0.40 mmol)and sodium triacetoxyborohydride (127 mg, 0.60 mmol) in DCE (5 mL) wasstirred for 24 h. The reaction mixture was diluted by DCM, filtered, andwashed with saturated aqueous NaHCO₃, water and brine. The DCM layerswere dried over Na₂SO₄, filtered and concentrated. The residue waspurified on preparative TLC (1000 micron) (developed by 100% EtOAc) toyield the final title compound as a free base. Its HCl salt (56.2 mg)was formed by treatment with 4 N HCl/dioxane. ESI-MS calc. forC₂₄H₃₂F₆N₂O: 478.24; Found: 479 (M+H).

EXAMPLE 223

Example 223 was prepared starting from Intermediate 8 andtrans-4-aminocyclohexanol as detailed in Example 222. LC-MS calc. ForC24H32F6N2O2: 494.24; Found: 495 (M+H).

EXAMPLE 224

Example 224 was prepared starting from Intermediate 8 andtrans-2-aminocyclohexanol as detailed in Example 222. LC-MS calc. ForC24H32F6N2O2: 494.24; Found: 495 (M+H).

EXAMPLE 226

Step A

To a solution of Intermediate 1 (22 g, 130 mmol) in benzene (140 mL),was added thionyl chloride (21 mL, 290 mmol). The reaction mixture wasstirred at 40° C. for 4.5 h and then the solvent and volatile materialswere evaporated. Distillation under vacuum yielded Intermediate chloride(Step A, Example 226) (7.186 g, 30%) (bp. 105-107° C./3 mmHg). ¹H-NMR(500MHz, CDCl₃): 2.82 (d, 1H), 2.50-2.56 (m, 1H), 2.3-2.42 (m, 4H),2.07-2.15 (m, 1H), 1.00-1.04 (m, 6H).Step B

A mixture of DCE/trimethyl orthoformate (1:1, 200 mL) was used to swellthe 4-(4-formyl-3-methoxy-phenoxy)butylryl AM resin (01-64-0209,NovaBiochem, 0.55 mmol/g, 13 g) for 2.45 h. After filtration, the resinwas suspended in DCE/trimethyl orthoformate (1:1, 180 mL) and the3,5-bis(trifluoromethyl)benzyl amine (22 g, 89 mmol) and sodiumtriacetoxyborohydride (22.7 g, 107 mmol) was added into the suspension.The suspension was left to spin for 48 h with periodic release of thepressure for the first 6 h. The resin was then filtered off, washed byMeOH (three times), DCM (three times), DMF (three times) and then washedagain by MeOH (three times), DCM (three times), DMF (three times). Itwas dried under vacuum and used in the next step (Step B, Example 226).Step C

To a suspension of the intermediate from the previous step (10.5 g, 5.25mmol) in DCM (100 mL), was added the intermediate from Step A, Example226 (4.95 g, 26.3 mmol), and DIEA (4.57 mL, 26.3 mmol) under nitrogen.The reaction was left to spin for 48 h. The resin was filtered, washedby DCM (12 times), dried under vacuum and used as obtained in the nextstep.Step D

The intermediate from Step C, Example 226 (100 mg, 0.05 mmol) wasswelled in 5% trimethyl orthoformate/DCE for 30 minutes and then thesolvent was drained off. After 5% trimethyl orthoformate/DCE (1.5 mL),2,3-dimethyl-cyclohexylamineamine (0.25 mmol) and sodiumtriacetoxyborohydride (110 mg, 0.50 mmol) was added, the suspension wasspun for 48 h and pressure was released every 15 minutes for 3 h. Theresin was then filtered out, washed by MeOH (twice), DCM (five times),DMF (five times) and then washed again by MeOH (twice), DMF (five times)and DCM (five times). The resin was dried under vacuum and mixed with25% TFA/DCM (2 mL), and spun for 1 h, was filtered, and washed twicewith DCM. The filtrate was evaporated to yield the final compound as aTFA salt (7.8 mg). LC-MS calc. For C26H36F6N2O calc.: 506.27; Found: 507(M+H).

EXAMPLE 227

Example 227 was prepared starting from the intermediate prepared in StepC, Example 226 and 2-methylcyclohexylamine as detailed in Example226-Step D. LC-MS calc. For C25H34F6N2O: 492.26; Found: 493 (M+H).

EXAMPLE 228

To a solution of the secondary amine described in Example 181 (60 mg,0.11 mmol) in DCM (4 mL), was added formalin (37% aqueous solution, 96mg, 1.1 mmol), molecular sieve (4 Å, 500 mg), and sodiumtriacetoxyborohydride (120 mg, 0.56 mmol). The reaction mixture wasstirred over 48 h, it was filtered, washing with DCM and MeOH. Saturatedaqueous NaHCO₃ was added and the mixture was heated at 60° C. for 1.5 h,and evaporated to remove the volatiles. The resulting mixture wasdiluted with water (10 mL) and extracted with DCM (four times). Theorganic portion was separated, washed with water, brine, dried overNa₂SO₄, filtered and concentrated. The residue was purified bypreparative TLC (1000 micron) (developed by 5% [aqueous NH₄OH/MeOH(1/9)]in DCM) to yield the final compound as a free base. Its HCl salt (47 mg)was formed by treatment with 4 N HCl/dioxane. LC-MS calc. forC27H36F6N2O3: 550.26; Found: 551 (M+H).

EXAMPLE 229

Example 229 was prepared starting from the intermediate prepared in StepC, Example 181 and 3-fluoro-5-trifluoromethylbenzylamine as detailed inExample 181. LC-MS for calc. C25H34F4N2O3: 486.25; Found: 487 (M+H).

EXAMPLE 230

Example 230 was prepared starting from Example 229 as detailed inExample 181. LC-MS for calc. C26H36F4N2O3: 500.27; Found: 501 (M+H).

EXAMPLE 231

Example 231 was prepared as detailed in Example 169 except thatbenzylchloroformate in Step A was replaced by methanesulfonyl chloride.LC-MS for. C21H27F6N3O4 for [M+H]⁺ calc. 532.16; Found: 532.30.

EXAMPLE 232

Example 232 was prepared as detailed in Example 169 except thatbenzylchloroformate in Step A was replaced by acetic anhydride. LC-MSfor. C22H27F6N2O3 for [M+H]⁺ calc. 496.20; Found: 496.20.

EXAMPLE 233

Step A

To a cooled (0° C.) solution of the hydrochloride salt of Intermediate 7(600 mg, 1.8 mmol) and triethylamine (370 μL, 1.8 mmol) in DCM (15 mL)was added di-tert butyl dicarbonate (470 mg, 2.2 mmol). The reaction wasallowed to warm to room temperature. After 4.5 h, the reaction wasdiluted with DCM and washed with saturated aqueous NaHCO₃ and brine. Theorganic layer was dried over MgSO₄, filtered and concentrated to give700 mg of product (93%). ESI-MS calc. for C13H15F31NO2: 401; found 346(M-tert-Butyl), 402 (M+H).Step B

The product from Step A (840 mg, 2.1 mmol) was combined withtetrakis(triphenylphosphine)palladium (150 mg, 0.13 mmol) and zinccyanide (170 mg, 1.5 mmol) in DMF (deoxygenated) and heated to 80° C.After 4 h, the reaction was cooled to room temperature and partitionedbetween EA and aqueous 2 N NH₄OH. The organic layer was washed twicewith aqueous 2 N NH₄OH, then brine, and dried over MgSO₄, filtered andconcentrated under reduced pressure. The product was purified by mediumpressure liquid chromatography (silica gel, 0-50% EA/hexanes) to give500 mg of product (80%). ESI-MS calc. for C14H15F3N2O2: 300; found 245(M-tert-Butyl).Step C

The product from Step B (500 mg, 1.7 mmol) was treated with hydrogenchloride (4 N in dioxane, 15 mL) and stirred at room temperature. After2 h the reaction was concentrated under reduced pressure to give 360 mgof a white solid (90%). ESI-Ms calc. for C9H7F3N2: 200; found 201 (M+H).Step D

Intermediate 17 (230 mg, 0.84 mmol) was combined with the product fromStep C (300 mg, 1.3 mmol), PyBrop (390 mg, 0.84 mmol), DMAP (61 mg, 0.5mmol), and triethylamine (180 μL, 1.2 mmol) in DCM (50 mL). After 72 hat room temperature the reaction was diluted with DCM, washed withsaturated aqueous NaHCO₃ and brine. The organic layer was dried overMgSO₄, filtered and concentrated under reduced pressure. The product waspurified by reverse phase HPLC (C18, 20-100% MeCN/H₂O) and converted toits hydrochloride salt by addition of 2 N HCl in ethyl ether to give 160mg of a white solid (43%). ESI-MS calc. for C24H32F3N3O2: 451; found 452(M+H).

While the invention has been described and illustrated with reference tocertain particular embodiments thereof, those skilled in the art willappreciate that various adaptations, changes, modifications,substitutions, deletions, or additions of procedures and protocols maybe made without departing from the spirit and scope of the invention.For example, effective dosages other than the particular dosages as setforth herein above may be applicable as a consequence of variations inthe responsiveness of the mammal being treated for any of theindications with the compounds of the invention indicated above.Likewise, the specific pharmacological responses observed may varyaccording to and depending upon the particular active compounds selectedor whether there are present pharmaceutical carriers, as well as thetype of formulation and mode of administration employed, and suchexpected variations or differences in the results are contemplated inaccordance with the objects and practices of the present invention. Itis intended, therefore, that the invention be defined by the scope ofthe claims which follow and that such claims be interpreted as broadlyas is reasonable.

1. A compound of the formula I:

wherein: X is selected from the group consisting of: —O—, —NR²⁰—, —S—,—SO—, —SO₂—, and —CR²¹R²²—, —NSO₂R²⁰—, —NCOR²⁰—, —NCO₂R²⁰—,—CR²¹CO₂R²⁰—, —CR²¹OCOR²⁰—, —CO—, —O—C(CH₃)₂—O—, where R²⁰ is selectedfrom: hydrogen, C₁₋₆ alkyl, benzyl, phenyl, C₃₋₆ cycloalkyl where thealkyl, phenyl, benzyl, and cycloalkyl groups can be unsubstituted orsubstituted with 1-3 substituents where the substituents areindependently selected from: halo, hydroxy, C₁₋₃alkyl, C₁₋₃alkoxy,—CO₂H, —CO₂—C₁₋₆ alkyl, and trifluoromethyl, where R²¹ and R²² areindependently selected from: hydrogen, hydroxy, C₁₋₆ alkyl,—O—C₁₋₆alkyl, benzyl, phenyl, C₃₋₆ cycloalkyl where the alkyl, phenyl,benzyl, and cycloalkyl groups can be unsubstituted or substituted with1-3 substituents where the substituents are independently selected from:halo, hydroxy, C₁₋₃alkyl, C₁₋₃alkoxy, —CO₂H, —CO₂—C₁₋₆ alkyl, andtrifluoromethyl; R¹ is selected from: —C₁₋₆alkyl,—C₀₋₆alkyl-O—C₁₋₆alkyl, —C₀₋₆alkyl-S—C₁₋₆alkyl,—C₀₋₆alkyl-SO₁₋₂—C₁₋₆alkyl, —C₀₋₆alkyl-SO₂—NR²⁶—C₁₋₆alkyl,—(C₀₋₆alkyl)-(C₃₋₇cycloalkyl)-(C₀₋₆alkyl), hydroxy, —CO₂R²⁰,heterocycle, —CN, —NR²OR²⁶, —NR²⁶SO₂R²⁰, —NR²⁶COR²¹, —OCOR²⁰, andphenyl, where R²⁶ is selected from: hydrogen, C₁₋₆ alkyl, benzyl,phenyl, C₃₋₆ cycloalkyl where the alkyl, phenyl, benzyl, and cycloalkylgroups can be unsubstituted or substituted with 1-3 substituents wherethe substituents are independently selected from: halo, hydroxy,C₁₋₃alkyl, C₁₋₃alkoxy, —CO₂H, —CO₂—C₁₋₁₆ alkyl, and trifluoromethylwhere the alkyl and the cycloalkyl are unsubstituted or substituted with1-7 substituents where the substituents are independently selected from:halo, hydroxy, —O—C₁₋₃alkyl, trifluoromethyl, C₁₋₃alkyl, —O—C₁₋₃alkyl,—CO₂R²⁰, —SO₂R²⁰, —NHCOCH₃, —NHSO₂CH₃, -heterocycle, ═O, —CN, and wherethe phenyl and heterocycle are unsubstituted or substituted with 1-3substituents where the substituents are independently selected from:halo, hydroxy, C₁₋₃alkyl, C₁₋₃alkoxy and trifluoromethyl; R² is selectedfrom: hydrogen, C₁₋₆alkyl, trifluoromethyl, trifluoromethoxy, chloro,bromo, and phenyl; R³ is selected from: hydrogen, hydroxy, halo,C₁₋₆alkyl, —O—C₁₋₆alkyl, —NR²⁰R²¹, —NR²⁰CO₂R²¹, —NR²⁰CONR²⁰R²¹,—NR²⁰—SO₂—NR²⁰R²¹, —NR²⁰—SO₂—R²¹, heterocycle, —CN, —CONR²⁰R²¹, —CO₂R²⁰,—NO₂, —S—R²⁰, —SO—R²⁰, —SO₂—R²⁰, and —SO₂—NR²⁰R²¹; R⁴ is selected from:hydrogen, C₁₋₆alkyl, trifluoromethyl, trifluoromethoxy, chloro, bromo,and phenyl; R⁵ is selected from: C₁₋₆alkyl substituted with 1-6 fluoroand optionally substituted with hydroxyl, —O—C₁₋₆alkyl substituted with1-6 fluoro, —CO—C₁₆alkyl substituted with 1-6 fluoro, —S—C₁₋₁₆alkyl,-pyridyl, fluoro, chloro, bromo, and phenyl; R⁶ is selected from:hydrogen, C₁₋₆alkyl, trifluoromethyl, trifluoromethoxy, chloro, bromo,and phenyl; R⁷ is selected from: hydrogen, C₁₋₆alkyl, andtrifluoromethyl; R⁸ is selected from: hydrogen, C₁₋₆alkyl, where alkylmay be unsubstituted or substituted with 1-6 substituents where thesubstituents are chosen from the group: fluoro, C₁₋₃alkoxy, hydroxy,—CO₂R²⁰, fluoro, —O—C₁₋₃alkyl, where alkyl may be unsubstituted orsubstituted with 1-3 fluoro, and C₃₋₆ cycloalkyl, —O—C₃₋₆cycloalkyl,hydroxy, —CO₂R²⁰, —OCOR²⁰, phenyl, or R⁷ and R⁸ may be joined togethervia a C₂₋₄alkyl or a C₀₋₂alkyl-O—C₁₋₃alkyl chain to form a 5-7 memberedring; R⁹ is selected from: hydrogen, C₁₋₆alkyl, where alkyl may beunsubstituted or substituted with 1-6 substituents where thesubstituents are chosen from the group: fluoro, C₁₋₃alkoxy, hydroxy,—CO₂R²⁰, —CO₂R²⁰, hydroxy, and —O—C₁₋₆alkyl, where alkyl may beunsubstituted or substituted with 1-6 substituents where thesubstituents are chosen from the group: fluoro, C₁₋₃alkoxy, hydroxy,—CO₂R²⁰, or R⁸ and R⁹ may be joined together by a C₁₋₄alkyl chain or aC₀₋₃alkyl-O—C₀₋₃alkyl chain to form a 3-6 membered ring; R¹⁰ is selectedfrom: hydrogen, and C₁₋₆alkyl, where alkyl may be unsubstituted orsubstituted with 1-6 fluoro, fluoro, —O—C₃₋₆cycloalkyl, and—O—C₁₋₃alkyl, where alkyl may be unsubstituted or substituted with 1-6fluoro, or R⁸ and R¹⁰ may be joined together by a C₁₋₃alkyl chain or asingle bond to form a 3-6 membered ring; where the alkyl areunsubstituted or substituted with 1-3 substituents where the substiuentsare independently selected from: halo, hydroxy, —CO₂R²⁰, C₁₋₃alkyl, andC₁₋₃alkoxy, or R⁸ and R¹⁰ may be joined together by aC₁₋₂alkyl-O—C₁₋₂alkyl chain to form a 6-8 membered ring, where the alkylare unsubstituted or substituted with 1-3 substituents where thesubstiuents are independently selected from: halo, hydroxy, —CO₂R²⁰,C₁₋₃alkyl, and C₁₋₃alkoxy, or R⁸ and R¹⁰ may be joined together by a—O—C₁₋₂alkyl-O— chain to form a 6-7 membered ring, where the alkyl areunsubstituted or substituted with 1-3 substituents where the substiuentsare independently selected from: halo, hydroxy, —CO₂R²⁰, C₁₋₃alkyl, andC₁₋₃alkoxy; R¹¹ is selected from: hydrogen, C₁₋₆alkyl, andtrifluoromethyl; R²⁷ and R²⁸ are independently selected from: ═O, whereR²⁷, R²⁸, or both, is oxygen and is connected via a double bond,hydrogen, phenyl, and C₁₋₆alkyl which may be substituted orunsubstituted with 1-6 of the following substituents: —COR¹¹, hydroxy,fluoro, chloro, —O—C₁₋₃alkyl; R²⁹, R³⁰, and R³¹ are independentlyselected from: hydrogen, methyl, hydroxyl, trifluoromethyl, methoxy, andtrifluoromethoxy; or R²⁹ and R⁹ are connected by a C₁₋₃alkyl bridge; mis selected from 0, 1, and 2; n is selected from 0, 1 and 2; the dashedline represents a single or a double bond; and pharmaceuticallyacceptable salts thereof and individual diastereomers thereof.
 2. Thecompound of claim 1 of the formula Ia:

and pharmaceutically acceptable salts and individual diastereomersthereof.
 3. The compound of claim 1 wherein: X is selected from thegroup consisting of: —O—, and —CH₂—.
 4. The compound of claim 1 whereinX is —O—.
 5. The compound of claim 1 wherein R¹ is selected from: (1)—C₁₋₆alkyl, which is unsubstituted or substituted with 1-6 substituentswhere the substituents are independently selected from: halo, hydroxy,—O—C₁₋₁₃alkyl, and trifluoromethyl, (2) —C₀₋₆alkyl-O—C₁₋₆alkyl-, whichis unsubstituted or substituted with 1-6 substituents where thesubstituents are independently selected from: halo, and trifluoromethyl,(3) —C₀₋₆alkyl-S—C₁₋₆alkyl-, which is unsubstituted or substituted with1-6 substituents where the substituents are independently selected from:halo, and trifluoromethyl, (4) —(C₃₋₅cycloalkyl)-(C₀₋₆alkyl), which isunsubstituted or substituted with 1-7 substituents where thesubstituents are independently selected from: halo, hydroxy,—O—C₁₋₃alkyl, and trifluoromethyl.
 6. The compound of claim 1 wherein R¹is C₁₋₆alkyl which is unsubstituted or substituted with 1-5 substituentswhere the substituents are independently selected from: hydroxy, andfluoro.
 7. The compound of claim 1 wherein: R¹ is selected from:isopropyl, —CH(OH)CH₃, and —CH₂CF₃.
 8. The compound of claim 1 wherein:R² is selected from: hydrogen, hydroxy, trifluoromethyl.
 9. The compoundof claim 1 wherein: R² is selected from: hydrogen, and hydroxy.
 10. Thecompound of claim 1 wherein: R³ is selected from: C₁₋₆alkylunsubstituted or substituted with 1-6 fluoro, fluoro, chloro, bromo. 11.The compound of claim 1 wherein: In the present invention it is morepreferred that R³ is selected from: trifluromethyl, cyclopropyl, fluoro.12. The compound of claim 1 wherein: R⁵ is selected from: C₁₋₆alkylunsubstituted or substituted with 1-6 fluoro, fluoro, chloro, bromo. 13.The compound of claim 1 wherein: R⁵ is selected from: trifluromethyl,cyclopropyl, and fluoro.
 14. The compound of claim 1 wherein: R⁵ istrifluoromethyl.
 15. The compound of claim 1 wherein R⁶ is hydrogen. 16.The compound of claim 1 wherein R⁷ is hydrogen.
 17. The compound ofclaim 1 wherein R⁸ is selected from: hydrogen, C₁₋₃alkyl, which isunsubstituted or substituted with 1-6 fluoro, —O—C₁₋₃alkyl, fluoro, andhydroxy.
 18. The compound of claim 1 wherein R⁸ is selected from:hydrogen, methyl, ethyl, trifluoromethyl, fluoro, and —O—CH₃.
 19. Thecompound of claim 1 wherein R⁹ is hydrogen and R¹⁰ is hydrogen.
 20. Thecompound of claim 1 wherein R⁸ and R¹⁰ are joined together by a —CH₂CH₂—chain or a —CH₂CH₂CH₂— chain to form a cyclopentyl ring or a cyclohexylring.
 21. The compound of claim 1 wherein R²⁷ is ═O, where R²⁷ is oxygenand is connected via a double bond.
 22. The compound of claim 1 whereinR⁹ and R²⁹ are joined together by a C₁₋₃alkyl chain to form a ring. 23.The compound of claim 1 wherein R²⁹ is hydrogen, R³⁰ is hydrogen, andR³¹ is hydrogen.
 24. A compound which is selected from the groupconsisting of the title compounds of the Examples, and pharmaceuticallyacceptable salts and individual diastereomers thereof.
 25. Apharmaceutical composition which comprises an inert carrier and acompound of claim
 1. 26. A method for modulation of chemokine receptoractivity in a mammal in need thereof which comprises the administrationof an effective amount of the compound of claim
 1. 27. A method fortreating, ameliorating or controlling an inflammatory orimmunoregulatory disorder or disease which comprises administering to apatient in need thereof an effective amount of the compound of claim 1.28. A method for reducing the risk of an inflammatory orimmunoregulatory disorder or disease which comprises administering to apatient in need thereof an effective amount of the compound of claim 1.29. A method for treating, ameliorating or controlling rheumatoidarthritis which comprises administering to a patient in need thereof aneffective amount of the compound of claim
 1. 30. A compound which isselected from the group consisting of:

and pharmaceutically acceptable salts thereof and individualdiastercomers thereof.
 31. A compound of the formula:

wherin R₇ is F or CF₃, and wherein R¹ is selected from:

and pharmaceutically acceptable salts thereof and individualdiastercomers thereof.
 32. A compound of the formula:

wherein R₂ is H or OH, wherein R₃ is F or CF₃, wherein R₄ is CF₃, Ph,OCF₃, Cl, or

and wherein R¹ is selected from:

and pharmaceutically acceptable salts thereof and individualdiastercomers thereof.
 33. A compound of the formula:

wherein R is selected from:

and pharmaceutically acceptable salts thereof and individualdiastercomers thereof.
 34. A compound of the formula:

wherein R is selected from:

and pharmaceutically acceptable salts thereof and individualdiastercomers thereof.
 35. A compound of the formula:

wherein R is selected from:

and pharmaceutically acceptable salts thereof and individualdiastercomers thereof.
 36. A compound of the formula:

wherein R is selected from:

and pharmaceutically acceptable salts thereof and individualdiastercomers thereof.